Parthenogenic activation of human oocytes for the production of human embryonic stem cells

ABSTRACT

Methods of producing human stem cells are disclosed for parthenogenetically activating human oocytes by manipulation of O 2  tension, including manipulation of Ca 2+  under high O 2  tension and contacting oocytes with serine threonine kinase inhibitors under low O 2  tension, isolating inner cell masses (ICMs) from the activated oocytes, and culturing the cells of the isolated ICMs under high O 2  tension. Moreover, methods are described for the production of stems cells from activated oocytes in the absence of non-human animal products, including the use of human feeder cells/products for culturing ICM/stem cells. Stem cells produced by the disclosed methods are also described.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. application Ser. No. 12/759,654 filed Apr. 13, 2010, now pending; which is a divisional application of U.S. application Ser. No. 11/505,260 filed Aug. 15, 2006, now U.S. Pat. No. 7,732,202; which claims the benefit under 35 USC §119(e) to U.S. Application Ser. No. 60/813,799 filed Jun. 14, 2006, now expired; to U.S. Application Ser. No. 60/758,443 filed Jan. 11, 2006, now expired; to U.S. Application Ser. No. 60/733,309 filed Nov. 2, 2005, now expired; and to U.S. Application Ser. No. 60/729,177 filed Oct. 21, 2005, now expired. The disclosure of each of the prior applications is considered part of and is incorporated by reference in the disclosure of this application.

which is a divisional application of U.S. application Ser. No. 11/505,260 filed Aug. 15, 2006, now U.S. Pat. No. 7,732,202; which claims the benefit under 35 USC §119(e) to U.S. Application Ser. No. 60/733,309 filed Nov. 2, 20015, now expired; to U.S. Application Ser. No. 60/758,443 filed Jan. 11, 2006, now expired; to U.S. Application Ser. No. 60/813,799 filed Jun. 14, 2006, now expired; and to U.S. Application Ser. No. 60/729,177 filed Oct. 21, 2005, now expired. The disclosure of each of the prior applications is considered part of and is incorporated by reference in the disclosure of this application.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates generally to embryonic stems cells, and more specifically to a process for obtaining human embryonic stem cells using parthenogenetically activated oocytes.

2. Background Information

Human embryonic stem cells (ES) cells are pluripotent cells that can differentiate into a large array of cell types. When injected into immune-deficient mice, embryonic stem cells form differentiated tumors (teratomas). However, embryonic stem into tissue types suitable for a patient's needs or the appropriate pharmacological test. Associated with this is a need for an efficient and reliable method of producing differentiated cells from embryonic stem cells.

Currently, human embryonic stem cells (hES) are derived from three sources: blastocysts remaining after infertility treatments and donated for research, blastocysts generated from donated gametes (oocytes and sperm), and the products of nuclear transfer (NT). Cadaveric fetal tissue is the only source of human embryonic germ cells (hEG). hES and hEG cells offer remarkable scientific and therapeutic possibilities, involving potential for generating more specialized cells or tissues. Ethical concerns about the sources of hES and hEG cells, however, and fears that use of NT for research could lead to use of NT to produce a human being, have fostered a great deal of public discussion and debate.

Parthenogenic activation of mammalian oocytes may be used as an alterative to fertilization by sperm/NT to prepare oocytes for embryonic stem cell generation. Parthenogenic activation is the production of embryonic cells, with or without eventual development into an adult, from a female gamete in the absence of any contribution from a male gamete.

Parthenogenetic activation of mammalian oocytes has been induced in a number of ways. Using an electrical stimulus to induce activation is of particular interest because electrofusion is part of the current nuclear transfer procedure. Parthenogenetic activation in vitro by electrical stimulation with electrofusion apparatus used for embryonic cell-oocyte membrane fusion has been reported.

Mouse oocytes have been activated by exposure to Ca⁺²-Mg⁺² free medium, medium containing hyaluronidase, exposure to ethanol, Ca⁺² ionophores or chelators, inhibitors of protein synthesis, and electrical stimulation. These procedures have led to high rates of parthenogenic activation and development of mouse oocytes, but did not activate and/or lead to a lower development rate of young bovine oocytes. Further, fertilization and parthenogenic activation of mouse oocytes is also dependent on post ovulatory aging.

Activation of bovine oocytes has been reported by ethanol, electrical stimulation, exposure to room temperature, and a combination of electrical stimulation and protein inhibition with cycloheximide. While these processes are thought to raise intracellular Ca⁺², they are most successful when the oocytes have been aged for more than 28 hours.

SUMMARY OF THE INVENTION

The present invention is based on the seminal discovery that certain conditions are optimal for parthenogenically activating human oocytes.

In one embodiment, a method of producing human stem cells is provided including parthenogenetically activating an oocyte, where activation includes contacting the oocyte with an ionophore at high oxygen (O₂) tension and contacting the oocyte with a serine-threonine kinase inhibitor under low O₂ tension, cultivating the activated oocyte at low O₂ tension until blastocyst formation, transferring the blastocyst to a layer of feeder cells, and culturing the transferred blastocyst under high O₂ tension, mechanically isolating an inner cell mass (ICM) from trophectoderm of the blastocyst, and culturing the cells of the ICM on a layer of feeder cells, where culturing the ICM cells is carried out under high O₂ tension. Preferably, the oocyte is human.

In a related aspect, low O₂ tension is maintained by incubation in a gas mixture environment comprising an O₂ concentration of about 2% O₂ to about 5% O₂, where the gas mixture environment further comprises about 5% CO₂ and about 90% nitrogen (N₂) to 93% N₂.

In another embodiment, a method of activating human metaphase II oocytes is provided including incubating human metaphase II oocytes in in vitro fertilization (IVF) media under high O₂ tension, activating by incubating the cells in IVF media comprising an ionophore under high O₂ tension, and subsequently incubating the cells in IVF media comprising a serine-threonine kinase inhibitor (STKI) under low O₂ tension, and incubating the STKI treated cells until blastocyst formation under low O₂ tension, where inner cell masses (ICM) obtained from the blastocyst produce culturable stem cells. High O₂ tension may be maintained by incubating the cells in a gas mixture environment having about 5% CO₂, about 20% O₂, and about 75% N₂.

In a related aspect, the O₂ tension for the incubating steps subsequent to activation is maintained by incubating the cells in a gas mixture environment comprising an O₂ concentration of about 2% O₂ to about 5% O₂, where the gas mixture environment further includes about 5% CO₂ and about 90% N₂ to about 93% N₂.

In another related aspect, the IVF media is essentially free of non-human products.

In a further related aspect, isolated oocytes prepared by the invention methods are provided, including isolated inner cell masses (ICM) prepared from such oocytes and corresponding stem cells isolated therefrom.

In another embodiment, human parthenogenic activation of mammalian oocytes resulting in embryogenic stem cells and their differentiated progeny is provided. Such cells and progeny are substantially isogenic to the oocyte donor, thus allowing for autologous transplantation of cells relative to the oocyte donor, and rejection by the oocyte donor's immune system is typically avoided.

In a related aspect, a cell bank of hES cell lines derived from parthenogenically activated oocytes is provided.

In one embodiment, a method for producing human stem cells from a cryopreserved oocyte or parthenote is provided, including microinjecting into the cytoplasm of the oocyte or parthenote a cryopreservation agent, freezing the oocyte or parthenote to a cryogenic temperature to cause it to enter a dormant state, storing the oocyte or parthenote in the dormant state, thawing the oocyte or parthenote, parthenogenically activating the oocyte, where the activation includes contacting the oocyte with an ionophore at high O₂ tension and contacting the oocyte with a serine-threonine kinase inhibitor under low O₂ tension, cultivating the parthenote or activated oocyte under low O₂ tension until blastocyst formation, isolating an inner cell mass (ICM) from the trophectoderm of the blastocyst, and culturing the cells of the ICM on a layer of feeder cells, where culturing is carried out under high O₂ tension.

In another embodiment, autologous stem cells derived from parthenogenetically activated oocytes from a human donor are provided. In one aspect, the stem cells possess a substantially identical haplotype as the donor cell. In a related aspect, stem cells are substantially identical genetically to the donor cell.

In one aspect, a stem cell is identified as a full sibling of the donor according to single nucleotide polymorphism (SNP) markers. In another aspect, a stem cell is genomically imprinted according to donor origin.

In one embodiment, a differentiated cell derived from a stem cell obtained from a parthenogenetically activated oocyte from a human donor is disclosed. In a related aspect, the differentiated cell includes, but is not limited to, a neuronal cell, a cardiac cell, a smooth muscle cell, a striated muscle cell, an endothelial cell, an osteoblast, an oligodendrocyte, a hematopoietic cell, an adipose cell, a stromal cell, a chondrocyte, an astrocyte, a dendritic cell, a keratinocyte, a pancreatic islet, a lymphoid precursor cell, a mast cell, a mesodermal cell, and an endodermal cell. In a further related aspect, the differentiated cell expresses one or more markers, including but not limited to, neurofiliment 68, NCAM, beta III-tubulin, GFAP, alpha-actinin, desmin, PECAM-1, VE-Cadherin, alpha-fetoprotein, or a combination thereof.

In another embodiment, a cell line comprising autologous stem cells is disclosed, where the stem cells are derived from parthenogenetically activated oocytes from a human donor. In one aspect, the cells do not express SSEA-1. In another aspect, the cells of the cell line give rise to ectodermal, mesodermal, and endodermal germ lines.

In one embodiment, a cell bank is disclosed including cryopreserved parthenotes, where the parthenotes are derived from parthenogenetically activated oocytes from one or more human donors. In a related aspect, the parthenotes have been cultivated under low O₂ tension until blastocyst formation.

In one embodiment, a cell bank is disclosed including cryopreserved autologous stem cells, where the stem cells are derived from parthenogenetically activated oocytes from one or more human donors.

In another embodiment, a method of treating a subject in need thereof, comprising administering a cellular composition comprising differentiated cells, wherein the differentiated cells are derived from a stem cell obtained from a parthenogenetically activated oocyte from a human donor. In one aspect, the differentiated cell is selected from the group consisting of a neuronal cell, cardiac cell, smooth muscle cell, striated muscle cell, endothelial cell, osteoblast, oligodendrocyte, hematopoietic cell, adipose cell, stromal cell, chondrocyte, astrocyte, dendritic cell, keratinocyte, pancreatic islet, lymphoid precursor cell, mast cell, mesodermal cell, and endodermal cell.

In a related aspect, the subject presents a disease selected from the group consisting of Parkinson's disease, Huntington's disease, Alzheimer's disease, ALS, spinal cord defects or injuries, multiple sclerosis, muscular dystrophy, cystic fibrosis, liver disease, diabetes, heart disease, retinal disease (such as macular degeneration and retinitis pigmentosa), cartilage defects or injuries, burns, foot ulcers, vascular disease, urinary tract disease, AIDS, and cancer.

In one embodiment, a method of generating cloned human embryonic stem cells is disclosed, including removing a first pronuclei from a previously fertilized human oocyte, transferring a second pro-nuclei into the enucleated oocyte, where the second pro-nuclei is derived from a donor oocyte or an oocyte from the mother of the donor, or a parthenogenetically activated oocyte, where the pro-nuclei of the oocyte has been replaced by the nucleus of a donor somatic cell prior to activation, and cultivating the resulting oocyte until blastocyst formation, where an inner cell mass from the blastocyst contains the embryonic stem cells.

Exemplary methods and compositions according to this invention are described in greater detail below.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A shows a micrograph of the surface marker expression of alkaline phosphatase for the parthenogenically derived hES cells.

FIG. 1B shows a micrograph of the expression for the surface marker Oct4.

FIG. 1C shows a micrograph of the expression for the surface marker SSEA-1.

FIG. 1D shows a micrograph of the expression for the surface marker SSEA-3.

FIG. 1E shows a micrograph of the expression for the surface marker SSEA-4.

FIG. 1F shows a micrograph of the expression for the surface marker TRA-1-60.

FIG. 1G shows a micrograph of the expression for the surface marker TRA-1-81.

FIG. 2A shows the analysis of telomerase activity for the parthenogenically derived hES cells. 500, 1000, and 10000 (units) of extract was used to perform the analysis. AH-heat treated test extract (negative control); positive control-telomerase positive cells; CHAPS-lysis buffer; TSR8-control template.

FIG. 2B shows a micrograph of embryoid body formation from parthenogenically derived hES cells, 9 day culture.

FIG. 2C shows a micrograph of embryoid body formation from parthenogenically derived hES cells, 10 day culture.

FIG. 2D illustrates the karyotype of parthenogenically derived hES cells.

FIG. 2E shows the results from DNA finger printing analysis of parthenogenically derived hES cells. 1-DNA from the blood of the oocyte donor; 2-DNA from the parthenogenic hES cells derived from the same donor; 3-DNA from human feeder fibroblasts.

FIG. 3 shows a Northern blot characterizing the expression of genes associated with genomic imprinting. DNA probes: SNRPN, Peg1_2, Peg1_A, H19, and GAPDH (as an internal control). NSF, neonatal skin fibroblasts; hES, human embryonic stem cell line derived from fertilized oocytes; 1, phESC-1; 2, phESC-3, 3, phESC-4, 4, phESC-5; 5, phESC-6; 6 phESC-7. NSF RT-, hES RT-, 1 RT-are negative controls.

FIG. 4 shows the differentiation of phESC into derivatives of all three germ layers. Ectoderm differentiation is presented by positive immunocytochemical staining for neuron specific markers 68 (A), NCAM (B), beta III-tubulin (C) and glial cell marker GFAP (D, M). Differentiated cells were positive for mesodermal markers: muscle specific alpha actinin (G) and desmin (J), endothelial markers PECAM-1 (E) and VE-Cadherin (F). Endoderm differentiation is presented by positive staining for alpha-fetoprotein (H, L). The phESC produce pigmented epithelial-like cells (I, K). Magnification (I)×100; (A-H, J-M), ×400.

FIG. 5 shows the characterization of phESC lines for specific markers. Undifferentiated colonies of phESC on human feeder layer cells (A-F), negative staining for SSEA-1 (G-L), expression of cell surface markers SSEA-3 (M-R), SSEA-4 (S-X). Magnification (A) to (E)×100; (F)×200; (G) to (X)×400. Alkaline phosphatase positive staining of phESC colonies on feeder cells (A-F), OCT-4 (G-L), TRA-1-60 (K-R) and TRA-1-81 (S-X). Magnification (A, B, O, R)×100; (C-F, M, S, X)×200; (G-L, N, P, Q, T-W)×400.

FIG. 6 demonstrates that phESC cells possess high levels of telomerase activity by comparison with positive control cells: “+”-extract from 500 cells; “−”-heat treated cell extract with inactivated telomerase; “Control+”-telomerase positive cell extract (applied with TRAPEZE Kit); “B”-CHAPS lysis buffer, primer-dimer/PCR contamination control; TSR8-telomerase quantitative control template (0.1 and 0.2 amole/μl); “M”-marker, DNA ladder.

FIG. 7 shows the G-banded karyotyping of phESC lines. The phESC-1 (A), phESC-3 (B), phESC-4 (C), phESC-5 (D) and phESC-6 (E) lines have normal 46, XX karyotype. The phESC-7 line has 47, XXX karyotype (F).

DETAILED DESCRIPTION OF THE INVENTION

Before the present composition, methods, and culturing methodologies are described, it is to be understood that this invention is not limited to particular compositions, methods, and experimental conditions described, as such compositions, methods, and conditions may vary. It is also to be understood that the terminology used herein is for purposes of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only in the appended claims.

As used in this specification and the appended claims, the singular forms “a”, “an”, and “the” include plural references unless the context clearly dictates otherwise. Thus, for example, references to “the method” includes one or more methods, and/or steps of the type described herein which will become apparent to those persons skilled in the art upon reading this disclosure and so forth.

“Differentiation” refers to a change that occurs in cells to cause those cells to assume certain specialized functions and to lose the ability to change into certain other specialized functional units. Cells capable of differentiation may be any of totipotent, pluripotent or multipotent cells. Differentiation may be partial or complete with respect to mature adult cells.

Gynogenesis refers to the production of an embryo containing a discernible trophectoderm and inner cell mass that results upon activation of a cell, such as an oocyte, or other embryonic cell type, containing mammalian DNA of all female origin, preferably human female origin, e.g., human or non-human primate oocyte DNA. Such female mammalian DNA may be genetically modified, e.g., by insertion, deletion or substitution of at least one DNA sequence, or may be unmodified. For example, the DNA may be modified by the insertion or deletion of desired coding sequences, or sequences that promote or inhibit embryogenesis. Typically, such an embryo will be obtained by in vitro activation of an oocyte that contains DNA of all female origin. Gynogenesis is inclusive of parthenogenesis which is defined below. It also includes activation methods where the spermatozoal DNA does not contribute to the DNA in the activated oocyte.

In a related aspect, oocytes are obtained from superovulating subjects prepared for IVF. “Superovulation” techniques, such as treatment of a female subject with hormones, used in IVF are designed to stimulate the ovaries to produce several eggs (oocytes) rather than the usual single egg as in a natural cycle.

The medications required to boost egg production may include, but are not limited to the following: Lupron (gonadotropin releasing hormone-agonist), Orgalutran, Antagon or Cetrotide (gonadotropin releasing hoi mone-antagonist), Follistim, Bravelle or Gonal-F (FSH, follicle stimulating hormone), Repronex (combination of FSH and LH, luteinizing hormone), and Pregnyl or Novarel (hCG, human chorionic gonadotropin).

In a related aspect, collection of eggs can be performed under transvaginal ultrasound guidance. To accomplish this, a needle is inserted (e.g., under IV sedation) through the vaginal wall into the ovaries using ultrasound to locate each follicle. The follicular fluid is drawn up into a test tube to obtain the eggs.

“Parthenogenesis” (“parthenogenically activated” and “parthenogenetically activated” is used interchangeably) the process by which activation of the oocyte occurs in the absence of sperm penetration, and refers to the development of an early stage embryo comprising trophectoderm and inner cell mass that is obtained by activation of an oocyte or embryonic cell, e.g., blastomere, comprising DNA of all female origin. In a related aspect, a “parthenote” refers to the resulting cell obtained by such activation. In another related aspect, “blastocyst” refers to a cleavage stage of a fertilized or activated oocyte comprising a hollow ball of cells made of outer trophoblast cells and an inner cell mass (ICM). In a further related aspect, “blastocyst formation” refers to the process, after oocyte fertilization or activation, where the oocyte is subsequently cultured in media for a time to enable it to develop into a hollow ball of cells made of outer trophoblast cells and ICM (e.g., 5 to 6 days).

In one embodiment, the process of creating cloned human embryonic stem cell line by parthenogenetically activated oocytes is disclosed. While pathogenesis is not an uncommon form of reproduction in nature, mammals are not known to be capable of this form of reproduction. However, a 10% rate of spontaneous parthenogenesis can be found in oocytes from females of the inbred mouse strain LT/Sv (Ozil and Huneau, Development (2001) 128:917-928; Vrana et al., Proc Natl Acad Sci USA (2003) 100 (Suppl 1):11911-11916; Berkowitz and Goldstein, New Eng J Med (1996) 335(23):1740-1748). Oocytes from placental mammals can be induced to undergo parthenogenesis in vitro; however, embryonic development is unsuccessful.

Following parthenogenic activation of mammalian oocytes and transfer of the activated oocyte into a surrogate mother, there is limited embryonic survival: ten days in mice; 21 days in sheep; 29 days in pigs; and 11.5 days in rabbits (Kure-bayashi et al., Theriogenology (2000) 53:1105-1119; Hagemann et al., Mol Reprod Dev (1998) 50:154-162; Surani and Barton, Science (1983) 222:10344036). The reason for this arrested development is likely due to genetic imprinting. It has been shown that maternal and paternal genomes are epigentically different and that both sets are required for successful embryonic development (Surani, Cell (1998) 93:309-312; Sasaki et al., (1992) 6:1843-1856). In parthenotes, all of the genetic material should be of maternal origin, a therefore should lack paternal imprinting. Paternal imprinting is thought to be responsible for extra-embryo tissue development, thus the development of trophoblastic tissue following fertilization of an enucleated oocyte (Stevens, Nature (1978) 276:266-267). In animals, therefore, enucleated zygotes may be useful for nuclear transfer with subsequent parthenogenic activation.

Mammalian parthenotes undergo only limited development with eventual death of the embryo. In Macac fascicularis, only 14 percent of oocytes in stage II metaphase following in vitro Parthenogenetic activation developed to the blastocyst stage following 8 days of culture (Monk, Genes Dev (1988) 2:921-925). Similarly, 12 percent of human oocytes that were parthenogenetically activated in vitro following nuclear transfer developed to the blastocyst state (Monk, 1988). In both cases, one stem cell line was created.

Embryos formed in spontaneously activated parthenotes in virgin females of the LT/Sv inbred mouse strain die within a few days. When nuclear transfer is performed from cells comprising the inner cell mass (ICM) of these embryos into fertilized enucleated C57BL/6j mouse oocytes, cloned mice with the LT/Sv genome are obtained (Kaufman et al., Nature (1977) 265:53-55). Thus, the use of a fertilized oocyte allows for full-term development of a parthenote. In one aspect, a fertilized enucleated human oocyte can be used to support development of a parthenogenetic embryo containing a donor's nuclei until the blastocyst stage.

In one embodiment, the pronuclei of a donor's oocyte or from the oocyte of the mother of a donor, following parthenogenetic activation, can be transferred into a fertilized human oocyte from which the male and female pronuclei have been extracted.

In another embodiment, a two stage process is disclosed for generating human stem cells including transferring the nucleus of a donor's somatic cell into a donor oocyte, where the oocyte is subsequently activated by parthenogenesis and transferring the pronuclei of the activated oocyte into a fertilized oocyte, where the male and female pronuclei have been extracted.

In another embodiment, the nucleus from a donor's somatic cell can be transferred into a fertilized enucleated human oocyte with subsequent parthenogenetic activation. The three embodiments above are illustrated by the following flow diagrams:

“Pluripotent cell” refers to a cell derived from an embryo produced by activation of a cell containing DNA of all female or male origin that can be maintained in vitro for prolonged, theoretically indefinite period of time in an undifferentiated state, that can give rise to different differentiated tissue types, i.e., ectoderm, mesoderm, and endoderm. The pluripotent state of the cells is preferably maintained by culturing inner cell mass or cells derived from the inner cell mass of an embryo produced by androgenetic or gynogenetic methods under appropriate conditions, for example, by culturing on a fibroblast feeder layer or another feeder layer or culture that includes leukemia inhibitory factor (LIF). The pluripotent state of such cultured cells can be confirmed by various methods, e.g., (i) confirming the expression of markers characteristic of pluripotent cells; (ii) production of chimeric animals that contain cells that express the genotype of the pluripotent cells; (iii) injection of cells into animals, e.g., SCID mice, with the production of different differentiated cell types in vivo; and (iv) observation of the differentiation of the cells (e.g., when cultured in the absence of feeder layer or LIF) into embryoid bodies and other differentiated cell types in vitro.

“Diploid cell” refers to a cell, e.g., an oocyte or blastomere, having a diploid DNA content of all male or female origin.

“Haploid cell” refers to a cell, e.g., an oocyte or blastomere, having a haploid DNA content, where the haploid DNA is of all male or female origin.

Activation refers to a process where a fertilized or unfertilized oocyte, for example, but not limited to, in metaphase II of meiosis, undergoes a process typically including separation of the chromatid pairs, extrusion of the second polar body, resulting in an oocyte having a haploid number of chromosomes, each with one chromatid. Activation includes methods whereby a cell containing DNA of all male or female origin is induced to develop into an embryo that has a discernible inner cell mass and trophectoderm, which is useful for producing pluripotent cells but which is itself is likely to be incapable of developing into a viable offspring. Activation may be carried out, for example, under one of the following conditions: (1) conditions that do not cause second polar body extrusion; (ii) conditions that cause polar body extrusion but where the polar body extrusion is inhibited; or (iii) conditions that inhibit first cell division of the haploid oocyte.

“Metaphase II” refers to a stage of cell development where the DNA content of a cell consists of a haploid number of chromosomes with each chromosome represented by two chromatids.

In one embodiment, metaphase II oocytes are activated by incubating oocytes under various O₂ tension gas environments. In a related aspect, the low O₂ tension gas environment is created by a gas mixture comprising an O₂ concentration of about 2%, 3%, 4%, or 5%. In a further related aspect, the gas mixture comprises about 5% CO₂. Further, the gas mixture comprises about 90% N₂, 91% N₂, or 93% N₂. This gas mixture is to be distinguished from 5% CO₂ air, which is approximately about 5% CO₂, 20% O₂, and 75% N₂.

“O₂ tension” refers to the partial pressure (pressure exerted by a single component of a gas mixture) of oxygen in a fluid (i.e., liquid or gas). Low tension is when the partial pressure of oxygen (pO₂) is low and high tension is when the pO₂ is high.

“Defined-medium conditions” refer to environments for culturing cells where the concentration of components therein required for optimal growth are detailed. For example, depending on the use of the cells (e.g., therapeutic applications), removing cells from conditions that contain xenogenic proteins is important; i.e., the culture conditions are animal-free conditions or free of non-human animal proteins. In a related aspect, “in vitro fertilization (IVF) media” refers to a nutrient system which contains chemically defined substances on or in which fertilized oocytes can be grown.

“Extracellular matrix (ECM) substrates” refer to a surface beneath cells which supports optimum growth. For example, such ECM substrates include, but are not limited to, Matrigel, laminin, gelatin, and fibronectin substrates. In a related aspect, such substrates may comprise collagen IV, entactin, heparin sulfate proteoglycan, to include various growth factors (e.g., bFGF, epidermal growth factor, insulin-like growth factor-1, platelet derived growth factor, nerve growth factor, and TGF-β-1).

“Embryo” refers to an embryo that results upon activation of a cell, e.g., oocyte or other embryonic cells containing DNA of all male or female origin, which optionally may be modified, that comprises a discernible trophectoderm and inner cell mass, which cannot give rise to a viable offspring and where the DNA is of all male or female origin. The inner cell mass or cells contained therein are useful for the production of pluripotent cells as defined previously.

“Inner cell mass (ICM)” refers to the inner portion of an embryo which gives rise to fetal tissues. Herein, these cells are used to provide a continuous source of pluripotent cells in vitro. Further, the ICM includes the inner portion of the embryo that results from androgenesis or gynogenesis, i.e., embryos that result upon activation of cells containing DNA of all male or female origin. Such DNA, for example, will be human DNA, e.g., human oocyte or spermatozoal DNA, which may or may not have been genetically modified.

“Trophectoderm” refers to another portion of early stage embryo which gives rise to placental tissues, including that tissue of an embryo that results from androgenesis or gynogenesis, i.e., embryos that result from activation of cells that contain DNA of all male or female origin, e.g., human ovarian or spermatozoan.

“Differentiated cell” refers to a non-embryonic cell that possesses a particular differentiated, i.e., non-embryonic, state. The three earliest differentiated cell types are endoderm, mesoderm, and ectoderm.

“Substantially identical” refers to a quality of sameness regarding a particular characteristic that is so close as to be essentially the same within the ability to measure difference (e.g., by HLA typing, SNP analysis, and the like).

“Histocompatible” refers to the extent to which an organism will tolerate a graft of a foreign tissue.

“Genomic imprinting” refers to the mechanism by which a number of genes throughout the genome are monoallelically expressed according to their parental origin.

“Homoplasmy,” including grammatical variations thereof, refers to the presence of the same type of the mitochondrial DNA (mtDNA) within a cell or individual.

“Heteroplasmy,” including grammatical variations thereof, refers to the presence of a mixture of more than one type of mitochondrial DNA (mtDNA) within a cell or individual.

“Uniparental” refers to one or more cells or individuals from which another arises and to which it remains subsidiary.

“Mechanically isolating” refers to the process of separating cell aggregates by physical forces. For example, such a process would exclude the use of enzymes (or other cell cleavage products) which might contain non-human materials.

In the native environment, immature oocytes (eggs) from the ovary undergo a process of maturation which results in the progression through meiosis to metaphase II of meiosis. The oocytes then arrest at metaphase II. In metaphase II, the DNA content of the cell consists of a haploid number of chromosomes, each represented by two chromatids.

Such oocytes may be maintained indefinitely by cryopreserving by, for example, but not limited to, microinjection with a sugar.

In one embodiment, a method for producing human stem cells from a cryopreserved oocyte or parthenote is provided, including microinjecting into the cytoplasm of the oocyte or parthenote a cryopreservation agent, freezing the oocyte or parthenote to a cryogenic temperature to cause it to enter a dormant state, storing the oocyte or parthenote in the dormant state, thawing the oocyte or parthenote, parthenogenically activating the oocyte under high O₂ tension in the presence or an ionophore followed by contacting the oocyte with a serine-threonine kinase inhibitor under low O₂ tension, culturing the activated oocyte or parthenote until blastocyst formation, isolating an inner cell mass (ICM) from the blastocyst, and culturing the cells of the ICM on a layer of human feeder cells, where culturing the ICM cells is carried out under high O₂ tension.

In one aspect, oocytes obtained as described are transferred to modified, isotonic IVF covered with embryo-tested mineral oil (Sigma), or any other suitable medium. If desired, the oocytes may be incubated with an extracellular sugar at the same concentration as the amount planned for microinjection. For example, to inject 0.1 M sugar, oocytes may be equilibrated in DMEM/F-12 with 0.1 M sugar. In one aspect, the cryopreservation agent comprises a lower Na⁺ concentration than standard DMEM (i.e., Na⁺ low media). In a related aspect, the cryopreservation agent comprises a higher K⁺ concentration than standard DMEM (i.e., K⁺ high). In a further related aspect, the cryopreservation agent comprises both a lower Na⁺ and higher K⁺ concentration than standard DMEM (i.e., Na⁺ low/K⁺ high media). In one aspect, the cryopreservation agent comprises an organic buffer, including but not limited to, HEPES. In another aspect, the cryopreservation agent comprises moieties that inhibit apoptotic protein (e.g., capases).

Alternatively, the oocytes may be optionally equilibrated with any other substantially non-permeable solute, such a NaCl, to decrease their cell volume prior to microinjection. This initial decrease in cell volume may result in a smaller final volume of the microinjected oocytes compared to oocytes not incubated in a hypertonic media prior to microinjection. This smaller final volume may minimize any potential adverse effect from the swelling of the oocytes. This general procedure for the preparation of cells for microinjection may also be used for other cell types (e.g., activated oocytes, hES cells, and the like).

The oocytes are then microinjected with a cryopreservation agent. Microinjection equipment and procedures are well characterized in the art and microinjection equipment known for use in injecting small molecules into cells may be used with the invention. In an exemplary microinjection step, oocytes can be microinjected at a pressure of 10 psi for 30 milliseconds. Another example of a standard microinjection technique is the method described by Nakayama and Yanagimachi (Nature Biotech. 16:639-642, 1998).

A cryopreservation agent useful in this process includes any chemical that has cryo-protective properties and is ordinarily non-permeable. In particular, the cryopreservation agent can include sugars either alone or mixed together with other traditional cryopreservation agents. Carbohydrate sugars such as trehalose, sucrose, fructose, and raffinose, may be microinjected to concentrations less than or equal to about 1.0 M, and more preferably, less than or equal to about 0.4 M. In one aspect, the concentration is between 0.05 and 0.20 M, inclusive. Additionally, an extracellular sugar or traditional cryopreservation agent may be added prior to storage. If the cells were incubated in a hypertonic solution prior to microinjection, the substantially non-permeable solute may be allowed to remain in the media after microinjection or may be removed from the media by washing the cells with media containing a lower concentration, or none, of this solute.

Certain sugars or polysaccharides which ordinarily do not permeate cell membranes because they are too large to pass through the membrane have superior physiochemical and biological properties for cryopreservation purposes. While these sugars ordinarily do not permeate cell membranes on their own, using the method as described, these ordinarily non-permeating sugars may be microinjected intracellularly to result in a beneficial effect.

Non-permeating sugars having a stabilizing or preserving effect on cells that are especially useful as the cryopreservation agent in the present method include sucrose, trehalose, fructose, dextran, and raffinose. Among these sugars, trehalose, a non-reducing disaccharide of glucose, has been shown to be exceptionally effective in stabilizing cell structures at low concentrations. The addition of extracellular glycolipids or glycoproteins may also stabilize the cell membrane.

Following the microinjection of the cryopreservation agent, the cells are prepared for storage. A variety of methods for freezing and/or drying may be employed to prepare the cells for storage. In particular, three approaches are described herein: vacuum or air drying, freeze drying, and freeze-thaw protocols. Drying processes have the advantage that the stabilized biological material may be transported and stored at ambient temperatures.

Typically, oocytes loaded with 1 to 2M DMSO are cooled at a very slow cooling rate (0.3 to 0.5° C./min) to an intermediate temperature (—60° C. to −80° C.) before plunging in liquid nitrogen for storage. The sample can then be stored at this temperature.

The suspended material can then be stored at cryopreservation temperatures, for example, by leaving the vials in liquid nitrogen (LN₂), for the desired amount of time.

Protocols for vacuum or air drying and for freeze drying proteins are well characterized in the art (Franks et al., “Materials Science and the Production of Shelf-Stable Biologicals,” BioPharm, October 1991, p. 39; Shalaev et al., “Changes in the Physical State of Model Mixtures during Freezing and Drying: Impact on Product Quality,” Cryobiol. 33, 14-26 (1996)) and such protocols may be used to prepare cell suspensions for storage with the method as described. In addition to air drying, other convective drying methods that may be used to remove water from cell suspensions include the convective flow of nitrogen or other gases.

An exemplary evaporative vacuum drying protocol useful with the method of the invention may include placing 20 μl each into wells on 12 well plates and vacuum drying for 2 hours at ambient temperature. Of course, other drying methods could be used, including drying the cells in vials. Cells prepared in this manner may be stored dry, and rehydrated by diluting in DMEM or any other suitable media.

A method of the invention using freeze drying to prepare the cells for storage begins with freezing the cell suspension. While methods of freezing known in the art may be employed, the simple plunge freezing method described herein for the freeze-thaw method may also be used for the freezing step in the freeze drying protocol.

After freezing, a two stage drying process may be employed. In the first stage, energy of sublimation is added to vaporize frozen water. Secondary drying is performed after the pure crystalline ice in the sample has been sublimated. Freeze dried cells can be stored and hydrated in the same manner as described above for vacuum drying. Viable cells may then be recovered.

After the recovery of cells from a frozen or dried state, any external cryopreservation agent may be optionally removed from the culture media. For example, the media may be diluted by the addition of the corresponding media with a lower concentration of cryopreservation agent. For example, the recovered cells may be incubated for approximately five minutes in media containing a lower concentration of sugar than that used for cell storage. For this incubation, the media may contain the same sugar that was used as the cryopreservation agent; a different cryopreservation agent, such as galactose; or any other substantially non-permeable solute. To minimize any osmotic shock induced by the decrease in the osmolarity of the media, the concentration of the extracellular cryopreservation agent may be slowly decreased by performing this dilution step multiple times, each time with a lower concentration of cryopreservation agent. These dilution steps may be repeated until there is no extracellular cryopreservation agent present or until the concentration of cryopreservation agent or the osmolarity of the media is reduced to a desired level.

The parthenogenetically activated oocytes, blastocysts, ICM, and autologous stem cells can be stored or “banked” in a manner that allows the cells to be revived as needed in the future. An aliquot of the parthenogenetically activated oocytes and autologous stem cells can be removed at any time, to be grown into cultures of many undifferentiated cells and then differentiated into a particular cell type or tissue type, and may then be used to treat a disease or to replace malfunctioning tissues in a subject. Since the cells are parthenogenetically derived from the donor, the cells can be stored so that an individual or close relative can have access to cells for an extended period of time.

In one embodiment, a cell bank is provided for storing parthenogenetically activated oocytes, blastocysts, ICM, and/or autologous stem cell samples. In another embodiment, methods for administering such a cell bank are provided. U.S. Published Patent Application No. 20030215942, which is incorporated by reference herein in its entirety, provides an example of a stem cell bank system.

Using methods such as those described above, the isolation and in vitro propagation of parthenogenetically activated oocytes, blastocysts, ICM, and autologous stem cell samples and their cryopreservation facilitates the establishment of a “bank” of transplantable human stem cells. Because it is possible to store smaller aliquots of cells, the banking procedure could take up a relatively small space. Therefore, the cells of many individuals could be stored or “banked” on a short term or long term basis, with relatively little expense.

In one embodiment, a portion of the sample is made available for testing, either before or after processing and storage.

This invention also provides methods of recording or indexing the parthenogenetically activated oocyte, blastocyst, ICM, and/or autologous stem cell samples so that when a sample needs to be located, it can be easily retrieved. Any indexing and retrieval system can be used to fulfill this purpose. Any suitable type of storage system can be used so that the parthenogenetically activated oocytes, blastocysts, ICM, and/or autologous stem cells can be stored. The samples can be designed to store individual samples, or can be designed to store hundreds, thousands, and even millions of different cell samples.

The stored parthenogenetically activated oocyte, blastocyst, ICM, and/or autologous stem cell samples can be indexed for reliable and accurate retrieval. For example, each sample can be marked with alphanumeric codes, bar codes, or any other method or combinations thereof. There may also be an accessible and readable listing of information enabling identification of each parthenogenetically activated oocyte, blastocyst, ICM, and/or autologous stem cell sample and its location in the bank and enabling identification of the source and/or type the cell sample, which is outside of the bank. This indexing system can be managed in any way known in the art, e.g., manually or non-manually, e.g. a computer and conventional software can be used.

In one embodiment, the cell samples are organized using an indexing system so that the sample will be available for the donor's use whenever needed. In other embodiments, the cell samples can be utilized by individuals related to the original donor. Once recorded into the indexing system, the cell sample can be made available for matching purposes, e.g., a matching program will identify an individual with matching type information and the individual will have the option of being provided the matching sample.

The storage banking system can comprise a system for storing a plurality of records associated with a plurality of individuals and a plurality of cell samples. Each record may contain type information, genotypic information or phenotypic information associated with the cell samples or specific individuals. In one embodiment, the system will include a cross-match table that matches types of the samples with types of individuals who wish to receive a sample.

In one embodiment, the database system stores information for each parthenogenetically activated oocyte, blastocyst, ICM, and/or autologous stem cell sample in the bank. Certain information is stored in association with each sample. The information may be associated with a particular donor, for example, an identification of the donor and the donor's medical history. For example, each sample may be HLA typed and the HLA type information may be stored in association with each sample. The information stored may also be availability information. The information stored with each sample is searchable and identifies the sample in such a way that it can be located and supplied to the client immediately.

Accordingly, embodiments of the invention utilize computer-based systems that contain information such as the donor, date of submission, type of cells submitted, types of cell surface markers present, genetic information relating to the donor, or other pertinent information, and storage details such as maintenance records and the location of the stored samples, and other useful information.

The term “a computer-based system” refers to the hardware, software, and any database used to store, search, and retrieve information about the stored cells. The computer-based system preferably includes the storage media described above, and a processor for accessing and manipulating the data. The hardware of the computer-based systems of this embodiment comprises a central processing unit (CPU) and a database. A skilled artisan can readily appreciate that any one of the currently available computer-based systems are suitable.

In one embodiment, the computer system includes a processor connected to a bus that is connected to a main memory (preferably implemented as RAM) and a variety of secondary storage devices, such as a hard drive and removable medium storage device. The removable medium storage device can represent, for example, a floppy disk drive, a DVD drive, an optical disk drive, a compact disk drive, a magnetic tape drive, etc. A removable storage medium, such as a floppy disk, a compact disk, a magnetic tape, etc. containing control logic and/or data recorded therein can be inserted into the removable storage device. The computer system includes appropriate software for reading the control logic and/or the data from the removable medium storage device once inserted in the removable medium storage device. Information relating to the parthenogenetically activated oocyte, blastocyst, ICM, and/or autologous stem cell can be stored in a well known manner in the main memory, any of the secondary storage devices, and/or a removable storage medium. Software for accessing and processing these data (such as search tools, compare tools, etc.) reside in main memory during execution.

As used herein, “a database” refers to memory that can store any useful information relating to the parthenogenetically activated oocyte and/or autologous stem cell collections and the donors.

The data relating to the stored parthenogenetically activated oocyte, blastocyst, ICM, and/or autologous stem cell can be stored and manipulated in a variety of data processor programs in a variety of formats. For example, the data can be stored as text in a word processing file, such as Microsoft WORD or WORDPERFECT, an ASCII file, an html file, or a pdf file in a variety of database programs familiar to those of skill in the art, such as DB2, SYBASE, or ORACLE.

A “search program” refers to one or more programs that are implemented on the computer-based system to search for details or compare information relating to the cryopreserved samples within a database. A “retrieval program” refers to one or more programs that can be implemented on the computer-based system to identify parameters of interest in the database. For example, a retrieval program can be used to find samples that fit a particular profile, samples having specific markers or DNA sequences, or to find the location of samples corresponding to particular individuals.

There is no upper limit on the number of cell samples that can be stored in one cell bank. In one embodiment, hundreds of products from different individuals will be stored at one bank or storage facility. In another embodiment, up to millions of products may be stored in one storage facility. A single storage facility may be used to store parthenogenetically activated oocyte and/or autologous stem cell samples, or multiple storage facilities may be used.

In some embodiments of the present invention, the storage facility may have a means for any method of organizing and indexing the stored cell samples, such as, for example, automated robotic retrieval mechanisms and cell sample manipulation mechanisms. The facility may include micromanipulation devices for processing cell samples. Known conventional technologies can be used for efficient storage and retrieval of the cell samples. Exemplary technologies include but are not limited to Machine Vision, Robotics, Automated Guided Vehicle System, Automated Storage and Retrieval Systems, Computer Integrated Manufacturing, Computer Aided Process Planning, Statistical Process Control, and the like.

The type information or other information associated with the individual in need of a sample may be recorded into a system that can be used to identify an appropriate matching product, such as, for example, a database system, an indexing system, and the like. Once recorded in the system, a match can be made between the type of the individual and a donor cell sample. In preferred embodiments, the donor sample is from the same individual as the individual in need of the sample. However, similar but not identical donor/recipient matches can also be used. The matching sample is available for the individual possessing the matching type identifier. In one embodiment of this invention, the individual's identification information is stored in connection with the cell sample. In some embodiments, the matching process occurs around the time of harvesting the sample, or can occur at any time during processing, storage, or when a need arises. Accordingly, in some embodiments of the invention, the matching process occurs before the individual is in actual need of the cell sample.

When the parthenogenetically activated oocyte, blastocyst, ICM, and/or autologous stem cell sample is needed by an individual, it may be retrieved and made available for research, transplantation or other purposes within minutes, if desired. The sample may also be further processed to prepare it for transplantation or other needs.

Normally, the oocyte is ovulated at this stage and fertilized by the sperm. The sperm initiates the completion of meiosis in a process called activation. During activation, the pairs of chromatids separate, the second polar body is extruded, and the oocyte retains a haploid number of chromosomes, each with one chromatid. The sperm contributes the other haploid complement of chromosomes to make a full diploid cell with single chromatids. The chromosomes then progress through DNA synthesis during the first cell cycle. These cells then develop into embryos.

By contrast, embryos described herein are developed by artificial activation of cells, typically mammalian oocytes or blastomeres containing DNA of all male or female origin. As discussed in the background of the invention, many methods have been reported in the literature for artificial activation of unfertilized oocytes. Such methods include physical methods, e.g., mechanical methods such as pricking, manipulation or oocytes in culture, thermal methods such as cooling and heating, repeated electric pulses, enzymatic treatments, such as trypsin, pronase, hyaluronidase, osmotic treatments, ionic treatments such as with divalent cations and calcium ionophores, such as ionomycin and A23187, the use of anesthetics such as ether, ethanol, tetracaine, lignocaine, procaine, phenothiazine, tranquilizers such as thioridazine, trifluoperazine, fluphenazine, chlorpromazine, the use of protein synthesis inhibitors such as cycloheximide, puromycin, the use of phosphorylation inhibitors, e.g., protein kinase inhibitors such as staurosporine, 2-aminopurine, sphingosine, and DMAP, combinations thereof, as well as other methods.

Such activation methods are well known in the art and are discussed U.S. Pat. No. 5,945,577, incorporated herein by reference.

In one embodiment, a human cell in metaphase II, typically an oocyte or blastomere comprising DNA of all male or female origin, is artificially activated for effecting artificial activation of oocytes.

In a related aspect, the activated cell, e.g., oocyte, which is diploid, is allowed to develop into an embryo that comprises a trophectoderm and an inner cell mass. This can be effected using known methods and culture media that facilitate blastocyst development.

After the gynogenetic embryos have been cultured to produce a discernable trophectoderm and inner cell mass, the cells of the inner cell mass are then used to produce the desired pluripotent cell lines. This can be accomplished by transferring cells derived from the inner cell mass or the entire inner cell mass onto a culture that inhibits differentiation. This can be effected by transferring the inner cell mass cells onto a feeder layer that inhibits differentiation, e.g., fibroblasts or epithelial cells, such as fibroblasts derived from postnatal human tissues, etc., or other cells that produce LIF. Other factors/components may be employed to provide appropriate culture conditions for maintaining cells in the undifferentiated state including, but not limited to, addition of conditioned media (Amit et al., Developmental Biol (2000) 227:271-278), bFGF and TGF-β1 (with or without LIF) (Amit et al., Biol Reprod (2004) 70:837-845), factors which activate the gp130/STAT3 pathway (Hoffman and Carpenter, Nature Biotech (2005) 23(6):699-708), factors which activate the PI3K/Akt, PKB pathway (Kim et al., FEBS Lett (2005) 579:534-540), factors that are members of the bone morphogenetic protein (BMP) super family (Hoffman and Carpenter (2005), supra), and factors which activate the canonical/β-catenin Wnt signaling pathway (e.g., GSK-3-specific inhibitor; Sato et al., Nat Med (2004) 10:55-63). In a related aspect, such factors may comprise culture conditions that include feeder cells and/or ECM substrates (Hoffman and Carpenter (2005), supra).

In one aspect, the inner cell mass cells are cultured on human postnatal foreskin or dermal fibroblast cells or other cells which produce leukemia inhibitory factor, or in the presence of leukemia inhibitory factor. In a related aspect, feeder cells are inactivated prior to seeding with the ICM. For example, the feeder cells can be mitotically inactivated using an antibiotic. In a related aspect, the antibiotic can be, but is not limited to, mitomycin C.

Culturing will be effected under conditions that maintain the cells in an undifferentiated, pluripotent state, for prolonged periods, theoretically indefinitely. In one embodiment, oocytes are parthenogenically activated with calcium ionophores under high O₂ tension followed by contacting the oocytes with a serine-threonine kinase inhibitor under low O₂ tension. The resulting ICM from the parthenogenically activated oocytes is cultured under high O₂ tension, where the cells, for example, are maintained using a gas mixture comprising 20% O₂. In one aspect, culturable refers to being capable of, or fit for, being cultivated. In a related aspect, ICM isolation is carried out mechanically after four days of blastocyst cultivation, where the cultivation is carried out on feeder cells. Such cultivation, for example, eliminates the need to use materials derived from animal sources, as would be the case for immunosurgery.

In a related aspect, culture media for the ICM is supplemented with non-animal sera, including but not limited to, human umbilical cord serum, where the serum is present in defined media (e.g., IVF, available from MediCult A/S, Denmark; Vitrolife, Sweden; or Zander IVF, Inc., Vero Beach, Fla.). In another aspect, the media and processes as provided are free of animal products. In a related aspect, animal products are those products, including serum, interferons, chemokines, cytokines, hormones, and growth factors, that are from non-human sources.

The pluripotent state of the cells produced by the present invention can be confirmed by various methods. For example, the cells can be tested for the presence or absence of characteristic ES cell markers. In the case of human ES cells, examples of such markers are identified supra, and include SSEA-4, SSEA-3, TRA-1-60, TRA-1-81 and OCT 4, and are known in the art.

Also, pluripotency can be continued by injecting the cells into a suitable animal, e.g., a SCID mouse, and observing the production of differentiated cells and tissues. Still another method of confirming pluripotency is using the subject pluripotent cells to generate chimeric animals and observing the contribution of the introduced cells to different cell types. Methods for producing chimeric animals are well known in the art and are described in U.S. Pat. No. 6,642,433, incorporated by reference herein.

Yet another method of confirming pluripotency is to observe ES cell differentiation into embryoid bodies and other differentiated cell types when cultured under conditions that favor differentiation (e.g., removal of fibroblast feeder layers). This method has been utilized and it has been confirmed that the subject pluripotent cells give rise to embryoid bodies and different differentiated cell types in tissue culture.

The resultant pluripotent cells and cell lines, preferably human pluripotent cells and cell lines, which are derived from DNA of entirely female original, have numerous therapeutic and diagnostic applications. Such pluripotent cells may be used for cell transplantation therapies or gene therapy (if genetically modified) in the treatment of numerous disease conditions.

In this regard, it is known that mouse embryonic stem (ES) cells are capable of differentiating into almost any cell type. Therefore, human pluripotent (ES) cells produced according to the invention should possess similar differentiation capacity. The pluripotent cells according to the invention will be induced to differentiate to obtain the desired cell types according to known methods. For example, human ES cells produced according to the invention may be induced to differentiate into hematopoietic stem cells, muscle cells, cardiac muscle cells, liver cells, islet cells, retinal cells, cartilage cells, epithelial cells, urinary tract cells, etc., by culturing such cells in differentiation medium and under conditions which provide for cell differentiation. Medium and methods which result in the differentiation of ES cells are known in the art as are suitable culturing conditions.

For example, Palacios et al, Proc. Natl. Acad. Sci., USA, 92:7530-7537 (1995) teach the production of hematopoietic stem cells from an embryonic cell line by subjecting stem cells to an induction procedure comprising initially culturing aggregates of such cells in a suspension culture medium lacking retinoic acid followed by culturing in the same medium containing retinoic acid, followed by transferal of cell aggregates to a substrate which provides for cell attachment.

Moreover, Pedersen, J. Reprod. Fertil. Dev., 6:543-552 (1994) is a review article which references numerous articles disclosing methods for in vitro differentiation of embryonic stem cells to produce various differentiated cell types including hematopoietic cells, muscle, cardiac muscle, nerve cells, among others.

Further, Bain et al, Dev. Biol., 168:342-357 (1995) teach in vitro differentiation of embryonic stem cells to produce neural cells which possess neuronal properties. These references are exemplary of reported methods for obtaining differentiated cells from embryonic or stem cells. These references and in particular the disclosures therein relating to methods for differentiating embryonic stem cells are incorporated by reference in their entirety herein. Thus, using known methods and culture medium, one skilled in the art may culture the subject ES cells, including genetically engineered or transgenic ES cells, to obtain desired differentiated cell types, e.g., neural cells, muscle cells, hematopoietic cells, etc. Pluripotent cells produced by the methods described herein may be used to obtain any desired differentiated cell type. Therapeutic usages of differentiated human cells are unparalleled. For example, human hematopoietic stem cells may be used in medical treatments requiring bone marrow transplantation. Such procedures are used to treat many diseases, e.g., late stage cancers such as ovarian cancer and leukemia, as well as diseases that compromise the immune system, such as AIDS. Hematopoietic stem cells can be obtained, e.g., by incorporating male or female DNA derived from a male or female cancer or AIDS patient with an enucleated oocyte, obtaining pluripotent cells as described above, and culturing such cells under conditions which favor differentiation, until hematopoietic stem cells are obtained. Such hematopoietic cells may be used in the treatment of diseases including cancer and AIDS.

Alternatively, the subject pluripotent cells may be used to treat a patient with a neurological disorder by culturing such cells under differentiation conditions that produce neural cell lines. Specific diseases treatable by transplantation of such human neural cells include, by way of example, Parkinson's disease, Alzheimer's disease, ALS and cerebral palsy, among others. In the specific case of Parkinson's disease, it has been demonstrated that transplanted fetal brain neural cells make the proper connections with surrounding cells and produce dopamine. This can result in long-term reversal of Parkinson's disease symptoms. In a related aspect, nerve precursors can be used to reanneal severed/damaged nerve fibers to restore movement after hand, leg, and spinal cord injuries.

One object of the subject invention is that it provides an essentially limitless supply of pluripotent, human cells that can be used to produce differentiated cells suitable for autologous transplantation for the oocyte donor. Human embryonic stem cells and their differentiated progeny derived from blastocysts remaining after infertility treatments, or created using NT, will likely be rejected by a recipient's immune system when used in allogenic cell transplantation therapy. Parthenogenically derived stem cells should result in differentiated cells that could alleviate the significant problem associated with current transplantation methods, i.e., rejection of the transplanted tissue which may occur because of host-vs-graft or graft-vs-host rejection relative to the oocyte donor. Conventionally, rejection is prevented or reduced by the administration of anti-rejection drugs such as cyclosporin. However, such drugs have significant adverse side-effects, e.g., immunosuppression, carcinogenic properties, as well as being very expensive. Cells produced by the methods as disclosed should eliminate, or at least greatly reduce, the need for anti-rejection drugs relative to the oocyte donor.

Another object of the subject invention is that it provides an essentially limitless supply of pluripotent, human cells that can be used to produce differentiated cells suitable for allogenic transplantation to members of the oocyte donor's family (e.g., siblings). The cells will be immunologically and genetically similar to those of the oocytes donor's direct family members and thus less likely to be rejected by the donor's family members.

Another object of this method is that parthenogenic activation of mammalian oocytes is a relatively simple procedure when compared to SCNT and results in the creation of stem cells with less cell manipulation.

Parthenogenic activation of mammalian oocytes has shown to be more efficient in the creation of stem cells than methods requiring mechanical manipulation of the oocyte (e.g., SCNT).

One drawback of SCNT is that subjects with deficient mitochondrial respiratory chain activity present phenotypes with striking similarities to abnormalities commonly encountered in SCNT fetuses and offspring (Hiendleder et al, Repro Fertil Dev (2005) 17(1-2):69-83). Cells normally contain only one type of mitochondrial DNA (mtDNA), termed homoplasmy, however, heteroplasmy does exist, usually as a combination of mutant and wild-type mt DNA molecules or form a combination of wild-type variants (Spikings et al., Hum Repro Update (2006) 12(4):401-415). As heteroplasmy can result in mitochondrial disease, various mechanisms exist to ensure maternal-only transmission. However, with the increasing use of protocols which bypass normal mechanisms for homoplasmy maintenance (e.g., cytoplasmic transfer (CT) and SCNT), perturbed mitochondrial function may be intrinsic to stem cells derived from these sources.

In one aspect, as the parthenotes are uniparental, the possibility of heteroplasmy is minimized.

Other diseases and conditions treatable by cell therapy include, by way of example, spinal cord injuries, multiple sclerosis, muscular dystrophy, diabetes, liver diseases Including acute diseases (viral hepatitis, drug overdoses (acetaminophen) and others), chronic diseases (chronic hepatitis and others (generally leading to cirrhosis)), heritable liver defects (hemophilia B, factor IX deficiency, bulirubin metabolism defects, urea cycle defects, lysosomal storage disease, al-antitrypsin deficiency and others), heart diseases, cartilage replacement, burns, foot ulcers, gastrointestinal diseases, vascular diseases, kidney disease, retinal disease, urinary tract disease, and aging related diseases and conditions.

This methodology can be used to replace defective genes, e.g., defective immune system genes, cystic fibrosis genes, or to introduce genes which result in the expression of therapeutically beneficial proteins such as growth factors, lymphokines, cytokines, enzymes, etc.

For example, the gene encoding brain derived growth factor may be introduced into human pluripotent cells produced according to the invention, the cells differentiated into neural cells and the cells transplanted into a Parkinson's patient to retard the loss of neural cells during such disease.

Also, the subject pluripotent human ES cells, may be used as an in vitro model of differentiation, in particular for the study of genes which are involved in the regulation of early development. Also, differentiated cell tissues and organs produced using the subject ES cells may be used in drug studies.

Further, the subject ES cells or differentiated cells derived therefrom may be used as nuclear donors for the production of other ES cells and cell colonies.

Still further, pluripotent cells obtained according to the present disclosure may be used to identify proteins and genes that are involved in embryogenesis. This can be effected, e.g., by differential expression, i.e., by comparing mRNAs that are expressed in pluripotent cells provided according to the invention to mRNAs that are expressed as these cells differentiate into different cell types, e.g., neural cells, myocardiocytes, other muscle cells, skin cells, etc. Thereby, it may be possible to determine what genes are involved in differentiation of specific cell types.

Further, ES cells and/or their differentiated progeny that have specific genetic defects, such as the genetic defect that leads to Duchene's Muscular Dystrophy, may be used as models to study the specific disease associated with the genetic defect.

Also, it is another object of the present disclosure to expose pluripotent cell lines produced according to the described methods to cocktails of different growth factors, at different concentrations and under different cell culture conditions such as cultured on different cell matrices or under different partial pressures of gases so as to identify conditions that induce the production and proliferation of desired differentiated cell types.

The following examples are intended to illustrate but not limit the invention.

Example 1 Production of Human Parthenogenic Embryogenic Stem Cells

Materials and Methods

Donors voluntarily donated oocytes, cumulous cells, and blood (for DNA analysis) with no financial payment. Donors signed comprehensive informed consent documents and were informed that all donated materials were to be used for research and not for reproductive purposes. Before ovarian stimulation, oocyte donors underwent medical examination for suitability according to FDA eligibility determination guidelines for donors of human cells, tissues, and cellular and tissue-based products (Food and Drug Administration. (Draft) Guidance for Industry: Eligibility Determination for Donors of Human Cells, Tissues, and Cellular and Tissue Based Products (HCT/Ps) dated May 2004) and order N 67 (Feb. 26, 2003) of Russian Public Health Ministry. It included X-ray, blood and urine analysis, and liver function test. Donors were also screened for syphilis, HIV, HBV, and HCV.

Oocytes were obtained using standard hormonal stimulation to produce superovulation in the subject donor. Each donor egg underwent ovarian stimulation by FSH from the 3rd to the 13th days of their menstrual cycle. A total of 1500 IU of FSh was given. From the 10th to the 14th day of the donor's menstrual cycle, gonadoliberin antagonist Orgalutran (Organon, Holland) was injected at 0.25 mg/day. From the 12th to the 14th day of the donor's menstrual cycle a daily injection of 75 IU FSH+75 IU LH (Menopur, Ferring GmbH, Germany) was given. If an ultrasound examination displayed follicles between 18 and 20 mm in diameter, a single 8000 IU dose of hGC (Choragon, Ferring GmbH, Germany) was administered on the 14th day of the donor's menstrual cycle. Trans-vaginal punction was performed 35 hours after hCG injection on approximately the 16th day. Follicular fluid was collected from the antral follicles of anesthetized donors by ultrasound-guided needle aspiration into sterile tubes.

Cumulus oocyte complexes (COCs) were picked from the follicular fluid, washed in Flushing Medium (MediCult) and then incubated in Universal IVF medium (MediCult, see Table 1) with a Liquid Paraffin (MediCult) overlay for 2 hours in a 20% O₂, 5% CO₂, at 37° C. humidified atmosphere.

TABLE 1 IVF media. COMPOSITION Calcium Chloride EDTA Glucose Human Serum Albumin Magnesium Sulfate Penicillin G Potassium Chloride Potassium di-Hydrogen Phosphate Sodium Bicarbonate Sodium Chloride Sodium Lactate Sodium Pyruvate Water

Before activation, cumulus-oocyte complexes (COCs) were treated with SynVitro Hyadase (MediCult, A/S, Denmark) to remove cumulus cells followed by incubation in Universal IVF medium with a paraffin overlay for 30 minutes.

From this point onward, the culture of oocytes and embryos was performed in a humidified atmosphere at 37° C. using O₂-reduced gas mixture (90% N₂+5% O₂+5% CO₂), with the exception of the ionomycin treatment. The oocytes were activated by incubation in 5 μM ionomycin for 5 minutes in a CO₂ incubator at 37° C. in a gas environment of 20% O₂, 5% CO₂, followed by culture with 1 mM 6-dimethylaminopurine (DMAP) for 4 hours in IVF medium, with paraffin overlay, in a gas environment of 90% N₂, 5% O₂, and 5% CO₂ at 37° C. The oocytes were then washed 3 times in IVF. Activation and cultivation were carried out in 4-well plates (Nunclon, A/S, Denmark) in 500 μl of medium overlaid with liquid paraffin oil (MediCult, A/S, Denmark).

Activated oocytes were cultivated in IVF medium in a gas environment comprising 5% O₂, 5% CO₂, and 90% N₂, and embryos generated from the activated oocytes were cultured in the same gas mixture.

Activated oocytes were allowed to incubate in IVF under the above conditions (i.e., low O₂ tension) until fully expanded blastocysts containing an inner cell mass (ICM) at a Blastocyst Scoring Modification of 1AA or 2AA (Shady Grove Fertility Center, Rockville, Md., and Georgia Reproductive Specialists, Atlanta, Ga.) was observed.

The zona pellucida was removed by 0.5% pronase (Sigma, St. Louis) treatment. The ICM from blastocysts was isolated by immuno-surgery where the blastocysts were incubated with horse antiserum to human spleen cells followed by exposure to guinea pig complement. Trophoectodern cells were removed from the ICM by gently pipetting the treated blastocysts.

For the derivation of ICM from whole blastocysts, the blastocysts were placed on a feeder layer in medium designed for culture of phESC (i.e., VitroHES™ media, e.g., DMEM/high glucose medium, VitroLife, Sweden) supplemented with 10% human umbilical cord blood serum, 5 ng/ml human recombinant LIF (Chemicon Int'l, Inc., Temecula, Calif.), 4 ng/ml recombinant human FGF (Chemicon Int'l, Inc., Temecula, Calif.) and penicillin-streptomycin (100 U/100 m)). When blastocysts attached and trophoplast cells spread, the ICM became visible. Through three to four days of additional culture, the ICM was isolated through mechanical slicing of the ICM from the trophoectoderm outgrowth using a finely drawn glass pipette. Further, the IMC cells were cultured on a feeder cell layer of mitotically inactivated post natal human dermal fibroblasts, in VirtroHES™ media (as formulated above) in a 96-well plate in 5% CO₂ and 20% O₂ at 37° C. This gas mixture was used to culture stem cells. Human fibroblast cultures were made using non-animal materials. Inactivation of fibroblasts was carried out using 10 μg/ml mitomycin C (Sigma, St. Louis, Mo.) for 3 hours.

In a separate method, immuno-surgery was performed by incubating blastocysts with horse antiserum to human spleen cells followed by exposure to rabbit complement. The trophectoderm cells were removed from the ICM through gentle pipetting of the treated blastocyts. Further culturing of the isolated ICMs was performed on a feeder layer of neonatal human skin fibroblasts (HSF) obtained from a genetically unrelated individual (with parental consent) derived using medium containing human umbilical cord blood serum. The HSF feeder layer was mitotically inactivated using mitomycin C.

The medium for the culture of HSF consisted of 90% DMEM (high glucose, with L-glutamaine (Invitrogen), 10% human umbilical cord blood serum and penicillin-streptomycin (100 U/100 mg) Invitrogen).

For the culture of ICM and phESC, VitroHES™ (Vitrolife) supplemented with 4 ng/ml hrbFGF, 5 ng/ml hrLIF and 10% human umbilical cord blood serum was used. The ICM was mechanically plated on a fresh feeder layer and cultured for three to four days. The first colony was mechanically cut and replated after five days of culture. All subsequent passages were made after five to six days in culture. For early passages, colonies were mechanically divided into clumps and replated. Further passing of phESC was performed with collagenase IV treatment and mechanical dissociation. The propagation of phESC was performed at 37° C., 5% CO₂ in a humidified atmosphere.

Oocyte Activation

From the initial donor, four oocytes were activated, and the activated oocytes were cultivated in IVF medium in a gas environment comprising 5% O₂, 5% CO₂, and 90% N₂ and followed over five (5) days. Table 2 shows the progress of maturation of the activated oocytes. Each oocyte was separated in a 4-well plate.

TABLE 2 Cultured Activated Oocytes.* Day 1 Day 2 Day 3 Day 5 N1 1 pronucleus (pn), 2 blastomers 4 bl equal, 1 morula, 1 polar body (pb) (bl) equal, fr-2% fr-15% fragmentation (fr)-0% N2 0 pn, 4 bl not equal, 5 bl not equal, 4 bl not equal, 1 pb fr-4% fr-20% fr-40% N3 1 pn, 2 bl not equal, 6 bl equal, early 1 pb ft-0% ft-0% blastocysts N4 1 pn, 4 bl equal, 4 bl equal, Fully expanded 1 pb fr-10% fr-20% blastocyst with good ICM IAA *Cells were incubated in M1 ™ media (MediCult) on the first day and M2 ™ media (MediCult) on days 2-5. Media was changed everyday. M1 ™ and M2 ™ contain human serum albumin, glucose and derived metabolites, physiological salts, essential amino acids, non-essential amino acids, vitamins, nucleotides, sodium bicarbonate, streptomycin (40 mg/l), penicillin (40.000 IU/l) and phenol red.

Inner cell masses were isolated from N4 and transferred to human fibroblast feeder cells as outlined above. N1 and N2 degenerated on Day 6. Further, on Day 6, N3 produced fully expanded blastocyst with ICM 2AB. N3 was then transferred to human fibroblast feeder cells on Day 6. ICM from N4 was unchanged. N3 was used to isolate stem cells.

ICM cells were cultivated in VitroHES™ medium in a gas environment comprising 5% CO₂, and 95% N₂ and followed over forty-five (45) days. Table 2a shows the progress of N3 ICM cell cultivation.

TABLE 2a Progress of N3-ICM Cultivation.* Day 3 ICM transplanted on fresh feeder cells. Day 8 Colony of cells divided mechanically into 6 pieces and cultivated in 3 wells of a 96-well plate-1st passage. Day 14 From five (5) colonies of 1st passage, cells were mechanically divided, and 20 colonies of a 2nd passage were cultivated in 3 wells of a 24-well plate. Day 20 Cells were plated in 35 mm dish-3rd passage. Day 24 Five (5) 35 mm dishes were seeded with cells-4th passage. One dish was divided chemically with 5% pronase (Sigma) at room temperature. Day 30 Twenty-five (25) 35 mm were seeded with cells-5th** passage. Day 34 6th** cell passage. Day 35 11 ampules were frozen from the 6th passage. Day 37 7th** cell passage. Day 44 12 ampules were frozen from the 7th passage. Day 45 8th cell passage. *Cells were grown on M2 ™ media (MediaCult). **These passages were made with pronase digestion.

Stem Cell Isolation.

From the oocytes from 5 donors, the use of MediCult media followed by a culture under reduced oxygen allowed for the production of 23 blastocysts on the fifth or sixth day of culture. Eleven of the blastocysts had visible ICMs (Table 3).

TABLE 3 Generation of Parthenotes and Parthenogenetic Embryonic Stem Cell Lines. Blastocysts derived Normally Without Donor Oocytes Oocytes activated Parthenotes With visible Lines Number harvested donated oocytes created ICM ICM generated 1 8 4 4 4 2 — phESC-1 immunosurgery 2 15 8 8 8 3 3 phESC-3 phESC-4 phESC-5 all from whole blastocysts 3 27 14  12¹ 11² 3 2 phESC-6 from whole blastocysts 4 22 11  10³ 10  2 3 phESC-7 from whole blastocysts 5 20  9⁴ 7 7 1 4 No cell line generated ¹two oocytes were not activated; ²one oocyte degenerated after activation; ³one oocyte was not activated; ⁴two oocytes were at metaphase stage I and were discarded.

These results indicate an approximate 57.5% success rate in the formation of blastocysts from parthenogenetically activated oocytes.

Immunohistochemical Staining

For immunostaining, hES cell colonies and phESC cells on feeder layers were seeded onto micro cover glass, washed twice with PBS and fixed with 100% methanol for 5 minutes at −20° C. Cells were washed twice with PBS+0.05% Tween-20 and permeabilized with PBS+0.1% Triton X-100 for 10 minutes at room temperature. After cell washing, non-specific binding was blocked by incubation with blocking solution (PBS+0.05% Tween-20+four percent goat serum plus three percent human umbilical cord blood serum) for 30 minutes at room temperature (RT). Monoclonal antibodies were diluted in blocking solution and used for one hour at RT: SSEA-1 (MAB4301) (1:30), SSEA-3 (MAB4303) (1:10), SSEA-4 (MAB4304) (1:50), OCT-4 (MAB4305) (1:30), TRA-1-60 (MAB4360) (1:50), and TRA-1-81 (MAB4381) (1:50) from Chemicon. After the cells were washed, secondary antibodies Alexa Fluor 546 (orange-fluorescent) and 488 (green-fluorescent) (Molecular Probes, Invitrogen) were diluted 1:1000 in PBS+0.05% Tween-20 and applied for one hour at RT. Cells were washed and nuclei were stained with DAPI (Sigma) 0.1 μg/ml in PBS+0.05% Tween-20 during ten minutes at RT. Cells were washed and mounted on slides with Mowiol (Calbiochem). Fluorescence images were visualized with a fluorescence microscope.

For the detection of mesodermal markers in three week old embryoid bodies or in contractile embryoid bodies, monoclonal mouse anti-desmina antibody anti-human alpha actinin antibody (Chemicon) as the muscle specific markers, and anti-human CD31/PECAM-1 antibody (R&D Systems), antihuman VE Cadherin (DC144) antibody (R&D Systems) as the endothelial markers were used.

For detection of the endodermal markers in embryoid bodies, monoclonal mouse anti-human alpha-fetoprotein antibody (R&D Systems) was used.

Alkaline Phosphatase and Telomerase Activity

Alkaline phosphatase and telomerase activity were performed according to the manufacturer's specifications with AP kit and TRAPEZE™ Kit (Chemicon).

Karyotyping

To analyse the karyotype, hES cells were treated with 10 μg/ml Demecolcine (Sigma) for two hours, harvested with 0.05% trypsin/EDTA (Invitrogen) and centrifuged at 700×rpm for three minutes. The pellet was resuspended in 5 ml of 0.56% KCl, and incubated for 15 minutes at RT. After repeated centrifugation, the supernatant was removed and cells were resuspended and fixed with 5 ml of an ice cold mixture of methanol/acetic acid (3:1) for five minutes at +4° C. The fixation of the cells was repeated twice, after that the cell suspension was placed onto microscope slides and the preparations were stained with Giemsa Modified Stain (Sigma). Metaphases from cells prepared in this manner were analyzed by a standard G-banding method. Quantity of 5/1000 metaphase spreads were revealed and 63 metaphases were analyzed.

Embryoid Body Formation

hES and phESC cell colonies were mechanically divided into clumps and placed in wells of a 24 well plate precoated with 1.5% agarose (Sigma) in medium containing 85% Knockout DMEM, 15% human umbilical cord blood serum, 1×MEM NEAA, 1 mM Glutamax, 0.055 mM β-mercaptoethanol, penicillin-streptomycin (50 U/50 mg), 4 ng/ml hrbFGF (all from Invitrogen, except serum). Human EBs were cultured for 14 days in suspension culture and placed on a culture dish to give outgrowth or cultivated in suspension for an additional week.

Neural differentiation was induced by the cultivation of two week old embryoid bodies attached to a culture dish surface over a period of a week in differentiation medium: DMEM/F12, B27, 2 mM Glutamax, penicillin-streptomycin (100 U/100 m) and 20 ng/ml hrbFGF (all from Invitrogen). Some embryoid bodies gave rise to differentiated cells with neural morphology, others were dissected and additionally cultured to produce neurospheres.

Rhythmically beating embryoid bodies appeared spontaneously following five days of culture after plating on an adhesive surface in the same medium as was used for embryoid body generation.

HLA Typing

Genomic DNA was extracted from donor blood, hES, phESC cells, and human newborn skin fibroblasts (NSFs) with Dynabeads DNA Direct Blood from Dynal (Invitrogen). HLA typing was performed by PCR with allele-specific sequencing primers (PCR-SSP, Protrans) according to the manufacturer's specifications. HLA class I genes (HLA A*,B*,Cw*) were typed with PROTRANS HLA A*B*Cw* defining A*01-A*80, B*07-B*83, Cw*01-Cw*18 regions. HLA class II genes (HLA DRB1*, DRB3*, DRB4*, DRB5*, DQA1*, DQB1*) were analyzed with PROTRANS HLA DRB1* defining DRB1*01-DRB1*16 (DR1-DR18), DRB3*, DRB4*, DRB5* regions and PROTRANS HLA DQB1* DQA1*defining DQB1*02-DQB1*06 (DQ2-DQ9), DQA1*0101-DQA1*0601 regions. PCR amplification was achieved: at 94° C. for 2 min; 10 cycles at 94° C. for 10 sec, 65° C. for 1 min; 20 cycles at 94° C. for 10 sec, 61° C. for 50 sec, 72° C. for 30 sec. Amplified products were detected in 2% agarose gel.

Affimetrix SNP Microarray Analysis

Genomic DNA was isolated from blood, cumulus cells, phESC and NSF by phenol/chloroform extraction method. These DNA samples obtained from four Caucasian subjects were genotyped with Affimetrix Mapping 50K Hind 240 Array (part of Affimetrix GeneChip Mapping 100K kit). Initially, the dataset contained 57,244 binary SNP markers. Since the number of markers is more than would be necessary to identify the equivalency of genomic samples and to study heterozygosity, 15 (chromosomes 1-15) out of 22 autosomal chromosomes were chosen. The shorter seven chromosomes were removed to reduce the chance that no marker, or only a single marker for a given chromosome, is selected during random sampling. The 1,459 markers were analyzed by Relcheck (version 0.67, Copyright © 2000 Karl W. Broman, Johns Hopkins University, Licensed under GNU General Public License version 2 (June 1991)).

Genomic Imprinting Analysis

Total nucleic acid was prepared as described Li et al. (J Biol Chem (2002) 277(16):13518-13527). RNA and DNA were extracted from cells using Tri-reagent (Sigma) or by using an RNA preparation kit from Qiagen (Valencia, Calif.).

Northern blots containing RNA from the various samples (see FIG. 3) were blotted onto filters by standard methods (See, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 1989, 2nd ed, Cold Spring Harbor Press). The Northern filter was hybridized with single stranded oligonucleotide probes that hybridized specifically to the mRNAs. The oligonucleotide probes were end-labeled with [γ³²P]ATP (Amersham Biosciences). The filters were subsequently washed three times for 10 min each with 0.2×SSC (1×SSC=0.15 M NaCl and 0.015 M sodium citrate) containing 0.1% SDS at 60° C. and analyzed by PhosphorImager (Molecular Dynamics). The sequences of the oligonucleotide probes were obtained from sequences based on the following Accession Nos.: NP002393 (Peg1_2 and Peg1_A; for these genes, human PEG1 is transcribed from two alternative promoters, resulting in the transcription of two isoforms, of which only one (isoform 1_2) is imprinted. Paternal expression isoform 1 occurs in conjunction with an unmethylated CpG island in exon 1 of the paternal allele, whereas the corresponding CpG island in the maternal gene (isoform 1_A) is fully methylated. See, e.g., Li et al. (2002), supra); CAG29346 (SNRPN); AF087017 (H19); NR_001564 (inactive X specific transcripts-XIST); and P04406 (GAPDH).

DNA Fingerprinting Analysis

Genomic DNA was isolated from blood, hES cells, and NSFs through a phenol/chloroform extraction, digested with HinfI restriction enzyme (Fermentas) and loaded in a 0.8% agarose gel. Following electrophoresis, denatured DNA was transferred to a nylon membrane (Hybond N, Amersham) by Southern blotting and hybridized with ³²P-labeled (CAC) 5 oligonucleotide probe. mData were analyzed after membrane exposition on X-ray film (Kodak XAR) using Cronex intensifying screens.

Monolocus PCR Genotyping

In order to determine allelic identities for minisatellite loci between blood donor DNA and stem cell DNA, 11 polymorphic sites ((1) 3′ Apolipoprotein B hypervariable minisatellite locus (3′ApoB); (2) D1S80 (PMCT118) hypervariable minisatellite locus (D1S80); (3) D6S366; (4) D16S359; (5) D7S820; (6) Human von Willebrand factor gene hypervariable minisatellite locus II (vWFII); (7) D13S317; (8) Human von Willebrand factor gene hypervariable microsatellite locus (vWA); (9) Human c-fms proto-oncogene for CFS-1 receptor gene microsatellite locus (CSF1PO); (10) Human thyroid peroxidase gene microsatellite locus (TPDX); and (11) Human tyrosine hydroxylase gene microsatellite locus (TH01)) were analyzed by PCR genotyping. Allele frequencies for known populations (i.e., Russian and Caucasian-American populations) determined for the above polymorphic sites were compared to allele frequencies of these sites in test samples (i.e., hES, NSF, and donor blood DNA). Chromosomal location, Genbank locus and locus definition, repeat sequence data, allelic ladder range, VNTR ladder size range, other known alleles, allele sizes, PCR protocols, and allele frequency results for the 11 minisatellite loci of the disclosed populations analyzed are provided below.

-   -   (1) 3′ Apolipoprotein B hypervariable minisatellite locus         (3′ApoB VNTR)

Chromosomal location: 2p23-p23

GenBank locus and locus definition: APOB, apolipoprotein B (including Ag(x) antigen) untranslated region

Repeat sequence 5′-3′:  (SEQ ID NO: 1) (TATAATTAAATATTTTATAATTAAAATATT)n

Allelic ladder size range (bases): 450+10+2 primer+links

VNTR ladder size range (# of repeats, according to Ludwig et al, 1989): 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52

Other known alleles (# of repeats): 25, 27, 28, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 54, 55

Promega K562 DNA® Allele sizes (# of repeats): 36/36

PCR Protocol:

Thermal cycler:DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:  Denaturation 94° C., 1′  Elongation and primer linking 60° C., 2′ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time The analysis may be done as described in Verbenko et al. (Apolipoprotein B 3′-VNTR polymorphism in Eastern European populations. Eur J Hum Gen (2003) 11(1):444-451). See Table 4.

TABLE 4 Allele Frequencies for Russian Populations Number of Alleles Allele Allele frequency observed 25 0.001 1 30 0.079 75 32 0.071 68 33 0.001 1 34 0.238 227 35 0.004 4 36 0.393 375 37 0.001 1 38 0.036 36 39 0.001 1 40 0.014 13 42 0.001 1 44 0.042 41 45 0.006 6 46 0.033 31 48 0.067 64 50 0.011 10 52 0.001 1 Homozygotes 94 Heterozygotes 333 Total samples 427

-   -   (2) D1S80 (pMCT118) hypervariable minisatellite locus (D1S80         VNTR)

Chromosomal location: 1p35-36

GenBank locus and locus definition: Human D1S80 and MCT118 gene

Repeat sequence 5′-3′:  (SEQ ID NO: 2) (GAAGACAGACCACAG)n 

Allelic ladder size range (bases): 387-762

VNTR ladder size range (# of repeats): 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 34, 35, 36, 37, 40, 41

Other known alleles (# of repeats): 13, 14, 15, 38, 39, >41

Promega K562 DNA® Allele sizes (# of repeats): 18/29

PCR Protocol:

Thermal cycler: DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:  Denaturation 94° C., 45″  Primer linking 60° C., 30″  Elongation 72° C., 45″ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time

The analysis may be done as described in Verbenko et al. (Allele frequencies for D1S80 (pMCT118) locus in some Eastern European populations. J Forensic Sci (2003) 48(1):207-208). See Table 5.

TABLE 5 Allele Frequencies for Russian Populations Number of Alleles Allele Allele frequency observed 18 0.280 33 20 0.017 2 21 0.009 1 22 0.042 5 23 0.017 2 24 0.390 46 25 0.017 2 26 0.025 3 28 0.068 8 29 0.009 1 30 0.034 4 31 0.059 7 33 0.017 2 34 0.008 1 36 0.008 1 Homozygotes 15 Heterozygotes 44 Total samples 59

-   -   (3) D6S366

Chromosomal location: 6q21-qter

GenBank locus and locus definition: NA

Allelic ladder size range (bases): 150-162

STR ladder size range (# of repeats): 12, 13, 15

Other known alleles (# of repeats): 10, 11, 14, 16, 17

Promega K562 DNA® Allele sizes (# of repeats): 13/14

PCR Protocol:

Thermal cycler: DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:  Denaturation 94° C., 1′  Elongation and primer linking 60° C., 2′ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time

The analysis may be done as described in Efremov et al. (An expert evaluation of molecular genetic individualizing systems based on the HUMvWFII and D6S366 tetranucleotide tandem repeats. Sud Med Ekspert (1998) 41(2):33-36). See Table 6.

TABLE 6 Allele Frequencies for Russian Populations Number of Alleles Allele Allele frequency observed 10 0.008 3 11 0.059 21 12 0.316 112 13 0.251 89 14 0.085 30 15 0.175 62 16 0.015 7 17 0.011 4 Total samples 177

-   -   (4) D16S539

Chromosomal location: 16q24-qter

GenBank locus and locus definition: NA

Repeat sequence 5′-3′:  (AGAT)n

Allelic ladder size range (bases): 264-304

STR ladder size range (# of repeats): 5, 8, 9, 10, 11, 12, 13, 14, 15

Promega K562 DNA® Allele sizes (# of repeats): 11/12

PCR Protocol:

Thermal cycler: DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:  Denaturation 94° C., 45″  Primer linking 64° C., 30″  Elongation 72° C., 30″ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time

The analysis has been done as described in GenePrint® STR Systems (Silver Stain Detection) Technical Manual No. D004. Promega Corporation, Madison, Wis. USA: 1993-2001. See Table 7.

TABLE 7 Allele Frequencies for Caucasian-Americans Allele Allele frequency Number of Alleles observed 6 0.000 0 7 0.000 0 8 0.026 11 9 0.107 45 10 0.079 33 11 0.319 134 12 0.269 113 13 0.167 70 14 0.031 13 15 0.002 1 Homozygotes 57 Heterozygotes 153 Total samples 210

-   -   (5) D7S820

Chromosomal location: 7q11.21-22

GenBank locus and locus definition: NA

Repeat sequence 5′-3′:  (AGAT)n

Allelic ladder size range (bases): 215-247

VNIK ladder size range (# of repeats): 6, 7, 8, 9, 10, 11, 12, 13, 14

Promega K562 DNA® Allele sizes (# of repeats): 9/11

PCR Protocol:

Thermal cycler: DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:   Denaturation 94° C., 45″   Primer linking 64° C., 30″   Elongation 72° C., 30″ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time

The analysis has been done as described in GenePrint® STR Systems (Silver Stain Detection) Technical Manual No. D004. Promega Corporation, Madison, Wis. USA: 1993-2001. See Table 8.

TABLE 8 Allele Frequencies for D7S820 in Different Populations Allele frequency for Number of Number of Caucasian- Alleles Allele frequency Alleles Allele Americans observed for Russians observed 6 0.002 1 0.0012 1 7 0.010 4 0.0087 7 8 0.155 65 0.1928 155 9 0.152 64 0.1480 119 10 0.295 124 0.2524 203 11 0.195 82 0.2040 164 12 0.121 51 0.1580 127 13 0.057 24 0.0299 24 14 0.012 5 0.0050 4 Homozygotes 43 92 Heterozygotes 167 310 Total samples 210 402

-   -   (6) Human von Willebrand factor gene hypervariable         microsatellite locus II (vWFII)

Chromosomal location: 12p13.3-12p13.2

GenBank locus and locus definition: HUMvWFII, Human von Willebrand factor gene

Repeat sequence 5′-3′:  (ATCT)n/(AGAT)n

Allelic ladder size range (bases): 154-178

STR ladder size range (# of repeats): 9, 11, 12, 13

Other known alleles (# of repeats): 8, 10, 14, 15

Promega K562 DNA® Allele sizes (# of repeats): 13/13

PCR Protocol:

Thermal cycler: DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:   Denaturation 94° C., 1′   Elongation and primer linking 60° C., 2′ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time

The analysis has been done as described in Efremov et al. (An expert evaluation of molecular genetic individualizing systems based on the HUMvWFII and D6S366 tetranucleotide tandem repeats. Sud Med Ekspert (1998) 41(2):33-36). See Table 9.

TABLE 9 Allele Frequencies for Russian Populations Number of Alleles Allele Allele frequency observed 9 0.082 37 10 0.088 40 11 0.392 177 12 0.296 134 13 0.069 31 14 0.058 26 15 0.015 7 Total samples 226

-   -   (7) D13S317

Chromosomal location: 13q22-q31

GenBank locus and locus definition: NA

Repeat sequence 5′-3′:  (AGAT)n

Allelic ladder size range (bases): 165-197

STR ladder size range (# of repeats): 8, 9, 10, 11, 12, 13, 14, 15

Other known alleles (# of repeats): 7

Promega K562 DNA® Allele sizes (# of repeats): 8/8

PCR Protocol:

Thermal cycler: DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:   Denaturation 94° C., 45″   Primer linking 64° C., 30″   Elongation 72° C., 30″ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time

The analysis has been done as described in GenePrint® STR Systems (Silver Stain Detection) Technical Manual No. D004. Promega Corporation, Madison, Wis. USA: 1993-2001. See Table 10.

TABLE 10 Allele Frequencies for D13S317 in Different Populations Number of Allele Number of Allele frequency for Alleles frequency Alleles Allele Caucasian-Americans observed for Russians observed 7 0.000 0 0 0 8 0.143 60 0.1393 112 9 0.052 22 0.0883 71 10 0.052 22 0.0684 55 11 0.305 128 0.3706 298 12 0.307 129 0.2040 164 13 0.083 35 0.0871 70 14 0.057 24 0.0423 34 15 0.000 0 0 0 Homozygotes 61 90 Heterozygotes 149 312 Total samples 210 402

-   -   (8) Human von Willebrand factor gene hypervariable         microsatellite locus (vWA)

Chromosomal location: 12p12pter

GenBank locus and locus definition: HUMVWFA31, Human von Willebrand factor gene

Repeat sequence 5′-3′:  (AGAT)n

Allelic ladder size range (bases): 139-167

STR ladder size range (# of repeats): 14, 16, 17, 18

Other known alleles (# of repeats): 11, 12, 13, 15, 19, 20, 21

Promega K562 DNA® Allele sizes (# of repeats): 16/16

PCR Protocol:

Thermal cycler: DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:   Denaturation 94° C., 1′   Elongation and primer linking 60° C., 2′ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time

The analysis has been done as described in GenePrint® STR Systems (Silver Stain Detection) Technical Manual No. D004. Promega Corporation, Madison, Wis. USA: 1993-2001. See Table 11.

TABLE 11 Allele Frequencies for HUMVWFA31 in Different Populations Number of Allele Number Allele frequency for Alleles frequency of Alleles Allele Caucasian-Americans observed for Russians observed 13 0.000 0 0.0025 2 14 0.131 56 0.0796 64 15 0.082 35 0.0920 74 16 0.211 90 0.2127 171 17 0.265 113 0.2836 228 18 0.202 86 0.2251 181 19 0.087 37 0.0833 67 20 0.021 9 0.0199 16 21 0.000 0 0.0012 1 Homozygotes 38 70 Heterozygotes 175 332 Total samples 213 402

-   -   (9) Human c-fms proto-oncogene for CSF-1 receptor gene         microsatellite locus (CSFIPO)

Chromosomal location: 5q33.3-34

GenBank locus and locus definition: HUMCSF1PO, Human c-fms proto-oncogene

Repeat sequence 5′-3 ′:  (AGAT)n

Allelic ladder size range (bases): 295-327

STR ladder size range (# of repeats): 7, 8, 9, 10, 11, 12, 13, 14, 15

Other known alleles (# of repeats): 6

Promega K562 DNA® Allele sizes (# of repeats): 9/10

PCR Protocol:

Thermal cycler: DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:   Denaturation 94° C., 45″   Primer linking 64° C., 30″   Elongation 72° C., 30″ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time

The analysis has been done as described in GenePrint® STR Systems (Silver Stain Detection) Technical Manual No. D004. Promega Corporation, Madison, Wis. USA: 1993-2001. See Table 12.

TABLE 12 Allele Frequencies for Caucasian-Americans Allele Allele frequency Number of Alleles observed 6 0.000 0 7 0.000 0 8 0.002 1 9 0.033 14 10 0.251 108 11 0.309 133 12 0.330 142 13 0.060 26 14 0.014 6 15 0.000 0 Homozygotes 47 Heterozygotes 168 Total samples 215

-   -   (10) Human thyroid peroxidase gene microsatellite locus (TPDX)

Chromosomal location: 2p25.1-pter

GenBank locus and locus definition: HUMTPOX, Human thyroid peroxidase gene

Repeat sequence 5′-3′:  (AATG)n

Allelic ladder size range (bases): 224-252

STR ladder size range (# of repeats): 6, 7, 8, 9, 10, 11, 12, 13

Other known alleles (# of repeats): none

Promega K562 DNA® Allele sizes (# of repeats): 8/9

PCR Protocol:

Thermal cycler: DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:   Denaturation 94° C., 45″   Primer linking 64° C., 30″   Elongation 72° C., 30″ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time

The analysis has been done as described in GenePrint® S′I′R Systems (Silver Stain Detection) Technical Manual No. D004. Promega Corporation, Madison, Wis. USA: 1993-2001. See Table 13.

TABLE 13 Allele Frequencies for Caucasian-Americans Allele Allele frequency Number of Alleles observed 6 0.002 1 7 0.000 0 8 0.528 227 9 0.093 40 10 0.056 24 11 0.284 122 12 0.037 16 13 0.000 0 Homozygotes 76 Heterozygotes 139 Total samples 215

-   -   (11) Human tyrosine hydroxylase gene microsatellite locus (TH01)

Chromosomal location: 5q33.3-34

GenBank locus and locus definition: HUMTHO1, Human tyrosine hydroxylase gene

Repeat sequence 5′-3′:  (AATG)n

Allelic ladder size range (bases): 179-203

STR ladder size range (# of repeats): 5, 6, 7, 8, 9, 10, 11

Other known alleles (# of repeats): 9.3

Promega K562 DNA® Allele sizes (# of repeats): 9.3/9.3

PCR Protocol:

Thermal cycler: DNA Technology Ltd., Russia Initial Incubation: 95° C., 2′ Cycling for 30 cycles:   Denaturation 94° C., 45″   Primer linking 64° C., 30″   Elongation 72° C., 30″ Extension step: 72° C., 5′ Hold step:  4° C., unlimited time

The analysis has been done as described in GenePrint® STR Systems (Silver Stain Detection) Technical Manual No. D004. Promega Corporation, Madison, Wis. USA: 1993-2001. See Table 14.

TABLE 14 Allele Frequencies for Caucasian-Americans Allele Allele frequency Number of Alleles observed 5 0.007 3 6 0.237 101 7 0.148 63 8 0.117 50 9 0.155 66 9.3 0.331 141 10 0.005 2 11 0.000 0 Homozygotes 50 Heterozygotes 163 Total samples 213

Results

The hES cells from this method display many features that are typical for embryonic stem cells: cytoplasmic lipid bodies, small cytoplasmic/nuclear ratio and clearly distinguishable nucleoli. The hES cell colonies display similar morphology to that reported previously for human embryonic stem cells derived after in vitro fertilization. The cells were immunoreactively positive for alkaline phosphatase (FIG. 1A), octamer-binding transcription factor 4 mRNA (Oct-4) (FIG. 1B), stage-specific embryonic antigen 1 (SSEA-1) (FIG. 1C), stage-specific embryonic antigen 3 (SSEA-3) (FIG. 1D), stage-specific embryonic antigen 4 (SSEA-4) (FIG. 1E), tumor rejection antigen 1-60 (TRA-1-60) (FIG. 1F), tumor rejection antigen 1-81 (TRA-1-81) (FIG. 1G), and negative for stage-specific embryonic antigen 1 (SSEA-1) (FIG. 1C), (which is positive for mouse embryonic stem cells, but not for human). Telomerase activity is often correlated with replicative immortality and is typically expressed in germ cells, cancer cells, and a variety of stem cells, including stem cells, but absent in most somatic cell types. The cells prepared by this method after three months in in vitro proliferation maintained their undifferentiated morphology and displayed high levels of telomerase activity (FIG. 2A). The pluripotency of the cells was investigated in vitro by embryoid body formation (FIG. 2B, 2C), G-banded karyotyping shows that cells have normal human 46XX karyotype (FIG. 2D).

DNA fingerprinting analysis was performed on the blood of the oocyte donor, on the ES cells, and on the HNSF feeder cells by Southern blotting and hybridization with a ³²P-labeled (CAC)s oligonucleotide probe (FIG. 2E), and monolocus polymerase chain reaction (PCR) with different locuses.

For monolocus PCR, genotyping revealed identical alleles for all loci (but one, D7S820) between blood (donor) DNA and OL1 DNA. See Table 15.

TABLE 15 Monolocus PCR genotyping. Chromosomal NN Locus definition location hES NSF Blood 1. 3′ApoB 2p24-p23 36/48 36/36 36/48 2. D1S80 1p35-36 18/24 22/31 18/24 3. D6S366 6q21-qter 13/15 17/17 13/15 4. D165359 16q24-qter  8/13 12/13 8/13 5. D7S820 7q11.21-22 11/11  9/10 10/11 6. vWFII 12p13.3-12p13.2 11/13  9/11 11/13 7. D135317 13q22-q31  9/12 11/12  9/12 8. vWA 12p12pter 14/18 17/18 14/18 9. CSF1PO 5q33.3-34 12/12 12/13 12/12 10. TPOX 2p25.1-pter  8/11  8/11  8/11 11. TH01 5q33.3-34 6/6   6/9.3 6/6

Heterozygosity (heterozygosis) of all heterozygous donor loci (but one, D7S820) was not changed in hES loci. Homozygosity (homozygosis) of D7S820 locus in hES DNA is a result of mutation (insertion of one AGAT monomer in microsatellite repeat) due to slipped-strand mispairing during DNA replication and DNA repair.

These results are in accordance with those obtained with multilocus DNA fingerprinting (when substantially identical fingerprint patterns for donor DNA and hES DNA were found).

FIG. 2E demonstrated heterozygosity of hES cells and their identity with the oocyte donor's blood, and there was no similarity between the hES cells and the feeder cells. The DNA profile of hES cell line was confirmed by PCR-based haplotype analysis using polymorphic genes within the MHC class I and class II. Total genomic DNA from the oocyte donor blood cells, from hES cells, and feeder HNSFs were genotyped and compared. The data demonstrated that hES cells and cells from donor blood were indistinguishable from each other and therefore should be considered autologous, and both distinguished from DNA of the feeder cells (Table 16).

TABLE 16 HLA Typing. MHC I MHC II HLA-A HLA-B HLA-C DRB1 DQB1 DQA1 pHES-1 A*01 B*15(63) Cw*04 DRB1*12 DQB1*06 DQA1*01 A*02 B*35 Cw*0708 DRB1*13 DQB1*03 DQA1*0505 Donor A*01 B*15(63) Cw*04 DRB1*12 DQB1*06 DQA1*01 A*02 B*35 Cw*0708 DRB1*13 DQB1*03 DQA1*0505 HNSF A*25 B*15(62) Cw*12 DRB1*04 DQB1*06 DQA1*01 A*32 B*18 Cw*12 DRB1*15 DQB1*03 DQA1*03

DNA fingerprinting and HLA typing analysis confirmed that the hES cells are heterozygous and contain the whole donor genetic material. These results coincide with data from parthenogenetic monkey stem cell lines (Vrana et al., Proc Natl Acad Sci USA (2003) 100(Suppl 1):11911-11916), and do not coincide with data from parthenogenetic mouse stem cell lines (Lin et al., Stem Cells (2003) 21:153-161), which contains half of the donor genetic material.

The phESC lines display a morphology expected in hES cells, forming colonies with tightly packed cells, prominent nucleoli and a small cytoplasm to nucleus ratio (FIG. 4). These cells express traditional hES markers SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, and OCT-4, and do not express SSEA-1, a positive marker for undifferentiated mouse embryonic stem cells (FIG. 4). The cells derived from all lines demonstrate high levels of alkaline phosphatase and telomerase activity (FIG. 5 and FIG. 6). G-banded karyotyping showed that phESC lines have a normal human 46,XX karyotype, with the exception of the phESC-7 line (FIG. 7). Approximately 91% of cells from the phESC-7 line have a 47,XXX karyotype and 9% of the cells have a 48,XXX,+6 karyotype. A different degree of X chromosome heteromorphism was observed in the lines; approximately 12% of the phESC-1 and phESC-6 lines; 42% for the phESC-5 line; and 70, 80, and 86% for the cell lines phESC7, phESC-3, and phESC-4, respectively (FIG. 7).

Comparative DNA profiling of was performed on all the phESC lines, the donor somatic cells and the feeder cells. These studies used Affimetrix SNP microarrays (Mapping 50K Hind 240 Arrays) to study chromosome changes and to confirm the genetic similarity of the phESC to the donor's somatic cells. All paired genotype relationships between phESC lines and their associated donor somatic cells were identified as “full siblings”, and all other combinations of pairs were identified as “unrelated”. Internal controls identified the paired genotype relationship between split cultures derived from the same phESC line as “monozygotic twins” (Table 17, Database S1).

TABLE 17 Database S1. Database S1 Identifying DNA samples from phESC and related donors putative inferred LOD LOD LOD LOD genotype 1 genotype 2 relationship relationship IBS 0 IBS 1 IBS 2 n_typed MZtwins par/off fullsibs halfsibs unrelated 1 2 unrelated unrelated 166 662 631 1459 −1503.03 −300.45 −23.15 −8.41 0 1 3 unrelated unrelated 241 616 602 1459 −1560.65 −434.85 −28.04 −12.22 0 1 4 unrelated unrelated 225 623 611 1459 −1535.94 −400.61 −31.39 −14.39 0 1 5 unrelated unrelated 225 623 611 1459 −1535.94 −400.61 −31.39 −14.39 0 1 6 unrelated unrelated 243 644 572 1459 −1642.35 −445.78 −31.74 −14.54 0 1 7 unrelated unrelated 252 638 569 1459 −1641.11 −453.5 −29.25 −12.86 0 1 8 unrelated unrelated 250 643 566 1459 −1656.02 −460.02 −32.86 −15.32 0 1 9 unrelated unrelated 219 657 583 1459 −1605.31 −382.39 −27.37 −11.58 0 1 10 unrelated unrelated 158 707 594 1459 −1591.43 −279.21 −26.37 −10 . . . 89 0 1 11 unrelated unrelated 193 668 598 1459 −1584.71 −354.76 −29.65 −13 . . . 31 0 1 12 unrelated unrelated 166 671 622 1459 −1523.1 −300.5 −30.53 −13 . . . 92 0 2 3 unrelated full sibs 0 282 1177 1459 −440.02 −146.3 0 −167.42 −363.63 2 4 unrelated unrelated 233 627 599 1459 −1569.66 −423.24 −28.24 −12.91 0 2 5 unrelated unrelated 233 627 599 1459 −1569.66 −423.24 −28.24 −12.91 0 2 6 unrelated unrelated 217 650 592 1459 −1584.75 −388.44 −22.62 −8.53 0 2 7 unrelated unrelated 243 650 566 1459 −1645.94 −437.91 −23.23 −8.72 0 2 8 unrelated unrelated 225 649 585 1459 −1603.18 −404.41 −27.04 −11.97 0 2 9 unrelated unrelated 210 639 610 1459 −1532.75 −360.46 −24.72 −9.89 0 2 10 unrelated unrelated 144 683 632 1459 −1491.18 −243.56 −16.82 −4.51 0 2 11 unrelated unrelated 172 680 607 1459 −1556.46 −310.03 −23.5 −9.7 0 2 12 unrelated unrelated 176 667 616 1459 −1538.57 −327.95 −27.31 −12 . . . 06 0 3 4 unrelated unrelated 336 457 666 1459 −1391.57 −599.92 −30.6 −14.62 0 3 5 unrelated unrelated 336 457 666 1459 −1391.57 −599.92 −30.6 −14.62 0 3 6 unrelated unrelated 322 482 655 1459 −1415.98 −571.23 −26.08 −11.86 0 3 7 unrelated unrelated 369 442 648 1459 −1432.05 −664.95 −27.39 −11.93 0 3 8 unrelated unrelated 334 483 642 1459 −1449.86 −597.75 −31.68 −15.14 0 3 9 unrelated unrelated 307 493 659 1459 −1395.19 −530.45 −24.56 −10 0 3 10 unrelated unrelated 215 623 621 1459 −1503.92 −364.97 −17.26 −4.43 0 3 11 unrelated unrelated 264 582 613 1459 −1531.91 −473.48 −28.41 −12 . . . 81 0 3 12 unrelated unrelated 254 595 610 1459 −1544.73 −460.57 −29.92 −13 . . . 88 0 4 5 unrelated MZ twins 0 0 1459 1459 0 −379.58 −45.47 −401.67 −677.74 4 6 unrelated unrelated 334 475 650 1459 −1436.59 −599.55 −32.73 −15.19 0 4 7 unrelated unrelated 365 439 655 1459 −1418.34 −656.01 −31.6 −14.56 0 4 8 unrelated unrelated 329 486 644 1459 −1450.75 −586.4 −32.06 −14.88 0 4 9 unrelated unrelated 332 466 661 1459 −1395.18 −590.12 −28.69 −12.94 0 4 10 unrelated unrelated 245 606 608 1459 −1542.32 −438.93 −28.75 −12 . . . 74 0 4 11 unrelated unrelated 273 569 617 1459 −1530.97 −492.84 −29.03 −12 . . . 34 0 4 12 unrelated full sibs 0 224 1235 1459 −326.17 −162.34 0 −183.44 −393.46 5 6 unrelated unrelated 334 475 650 1459 −1436.59 −599.55 −32.73 −15.19 0 5 7 unrelated unrelated 365 439 655 1459 −1418.34 −656.01 −31.6 −14.56 0 5 8 unrelated unrelated 329 486 644 1459 −1450.75 −586.4 −32.06 −14.88 0 5 9 unrelated unrelated 332 466 661 1459 −1395.18 −590.12 −28.69 −12.94 0 5 10 unrelated unrelated 245 606 608 1459 −1542.32 −438.93 −28.75 −12 . . . 74 0 5 11 unrelated unrelated 273 569 617 1459 −1530.97 −492.84 −29.03 −12 . . . 34 0 5 12 unrelated full sibs 0 224 1235 1459 −326.17 −162.34 0 −183.44 −393.46 6 7 unrelated full sibs 45 176 1238 1459 −277.78 −217.21 0 −165.72 −390.62 6 8 unrelated full sibs 44 187 1228 1459 −289.8 −201.32 0 −153.75 −365.51 6 9 unrelated unrelated 333 481 645 1459 −1436.5 −595.4 −30.3 −13.77 0 6 10 unrelated unrelated 240 601 618 1459 −1518.17 −425.03 −27.11 −11 . . . 53 0 6 11 unrelated full sibs 0 164 1295 1459 −209.27 −191.66 0 −213.25 −440.56 6 12 unrelated unrelated 234 615 610 1459 −1547.15 −416.14 −30.21 −13 . . . 64 0 7 8 unrelated full sibs 38 225 1196 1459 −326.62 −150.16 0 −121.55 −334.09 7 9 unrelated unrelated 359 473 627 1459 −1479.28 −642.41 −30.61 −14.47 0 7 10 unrelated unrelated 252 623 584 1459 −1598.35 −443.81 −28.88 −13 . . . 09 0 7 11 unrelated full sibs 0 230 1229 1459 −318.49 −137.93 0 −159.55 −389.58 7 12 unrelated unrelated 265 583 611 1459 −1539.33 −472.91 −30.55 −13 . . . 87 0 8 9 unrelated unrelated 347 480 632 1459 −1472.41 −625.68 −30.93 −14.31 0 8 10 unrelated unrelated 244 614 601 1459 −1561.3 −434 −28.07 −12 . . . 37 0 8 11 unrelated full sibs 0 175 1284 1459 −223.73 −178.56 0 −200.12 −428.04 8 12 unrelated unrelated 236 610 613 1459 −1539.08 −417.14 −29.32 −13 . . . 14 0 9 10 unrelated full sibs 0 228 1231 1459 −315.15 −152.88 0 −174.27 −392.91 9 11 unrelated unrelated 269 567 623 1459 −1502.69 −479.57 −28.47 −12 . . . 55 0 9 12 unrelated unrelated 245 612 602 1459 −1557.25 −438.53 −26.07 −11 . . . 15 0 10 11 unrelated unrelated 187 635 637 1459 −1478.7 −328.06 −25.52 −10.6 0 10 12 unrelated unrelated 181 662 616 1459 −1534.36 −329 −25.2 −10.6 0 DNA samples were numbered as follows: 1-human neonatal skin fibroblasts; 2-phESC-7 line donor; 3-phESC-7 line; 4-phESC-1 line; 5-phESC-1 line; 6-phESC-3 line; 7-phESC-4 line; 8-phESC-5 line; 9-phESC-6 line; 10-phESC-6 line donor; 11-phESC-3 to phESC-5 lines donor; and 12-phESC-1 line donor. The result shows that only one pair (sample 4-5), has been identified as monozygotic (MZ) twins. Ten other pairs (samples 2-3, 4-12, 5-12, 6-7, 6-11, 7-8, 7-11, 8-11, 9-10) have been identified as full siblings, and all the other combination of pairs have been identified as unrelated. The IBS columns in the output display the number of markers at which the pair are both typed and share 0, 1, or 2 alleles identical by state (For MZ twins under ideal conditions of no genotyping errors, all markers must be placed under IBS = 2). The output does not display P (observed markers | given relationship) directly, but it displays LOD score − log₁₀ {P(observed markers | putative relationship/P(observed markers | relationship for which maximum likelihood was obtained and thus the call was made)} as a measure of similarity. The smaller the LOD score is, the less likely the putative relationship between two samples it.

Comparative analysis of 1,459 SNP markers revealed phESC heterozygosity and showed that changes had occurred in the phESC cell genotype in comparison to the related donor somatic cell genotype. Some segments of the somatic cell genome that had formerly been heterozygous became homozygous in the related phESC line genome. This heterozygous to homozygous pattern occurred in 11-15% of the phESC-1, PhESC-3, phESC-4, phESC-5 and phESC-6 lines, and was 19% for the phESC7 line (Database S2). Moreover, genetic differences were observed between the phESC and phESC-5 lines that had been derived from the same oocyte donor (Table 18, Database S2).

TABLE 18 Database S2. Database S2 Heterozygosity of phESC (Abbreviated as “pC”) Lines chromo- RS ID basepair Freq A in pC-1 N3-5 pC-6 pC-7 some SNP ID (dbSNP) basepair (Mb) Caucasian pC-1 donor pC-3 pC-4 pC-5 donor pC-6 donor pC-7 donor 1 SNP_A- rs10752719 3744122 3.744122 0.436 AB AB BB AB AB AB BB BB BB BB 1697748 1 SNP_A- rs806104 5977200 5.9772 0.631 AB AB AA AB AB AB AB AB AA AA 1743594 1 SNP_A- rs301791 8402638 8.402638 0.274 AB AB BB BB BB BB AB AB AA AB 1687843 1 SNP_A- rs1474868 11978430 11.97843 0.333 BB BB AA AA AA AA AA AA AA AA 1647681 1 SNP_A- rs1417144 14211391 14.211391 0.548 AA AA AB AB AB AB AA AA AB AB 1673737 1 SNP_A- rs860379 18414756 18.414756 0.25 BB BB BB BB BB BB BB BB BB BB 1747116 1 SNP_A- rs10492997 19514677 19.514677 0.631 AA AA BB BB BB BB AB AB AA AA 1662223 1 SNP_A- rs559346 28077956 28.077956 0.583 BB AB AB AB AB AB AB AB AA AB 1662225 1 SNP_A- rs4949455 31783886 31.783886 0.382 AA AB AB BB AB AB AB AB AA AA 1646469 1 SNP_A- rs6661190 33872782 33.872782 0.274 BB BB BB BB BB BB BB BB BB AB 1695076 1 SNP_A- rs4653029 34558585 34.558585 0.274 BB BB BB BB BB BB BB BB BB BB 1679571 1 SNP_A- rs7531479 36798680 36.79868 0.714 AA AA AA AA AA AA AA AA AA AA 1675060 1 SNP_A- rs1010805 37858235 37.858235 0.738 AA AA AB AA BB AB AA AA BB BB 1753902 1 SNP_A- rs6693076 39972196 39.972196 0.441 AA AA AB AA BB AB AB AB BB BB 1691977 1 SNP_A- rs407752 40472948 40.472948 0.333 BB BB AB AA BB AB BB BB BB BB 1723259 1 SNP_A- rs7515340 41055964 41.055964 0.381 AB AB AA AA AA AA AA AA BB BB 1692103 1 SNP_A- rs4660575 41902429 41.902429 0.429 BB BB BB BB BB BB AA AA BB AB 1696731 1 SNP_A- rs1408412 42787470 42.78747 0.679 AA AA AA AA AA AA AA AA AA AA 1701070 1 SNP_A- rs1771551 45552736 45.552736 0.738 AA AA AA AA AA AA AA AA BB AB 1729559 1 SNP_A- rs2245122 47358015 47.358015 0.598 AA AA AA AA AA AA BB BB BB AB 1670587 1 SNP_A- rs1875645 50501900 50.5019 0.524 AB AB BB BB BB BB BB BB BB BB 1711898 1 SNP_A- rs625643 54349188 54.349188 0.25 BB BB BB BB BB BB BB BB BB BB 1645411 1 SNP_A- rs10493206 56531172 56.531172 0.488 AA AA BB BB BB BB AB AB AA AB 1718210 1 SNP_A- rs1831870 57339224 57.339224 0.524 AB AB AB AA BB AB BB BB BB AB 1752670 1 SNP_A- rs852766 57998529 57.998529 0.564 AA AA AA AA AA AA AA AA BB AB 1669308 1 SNP_A- rs1969772 58917123 58.917123 0.738 AA AA AA AA AA AA AB AB AA AB 1681141 1 SNP_A- rs10489908 61576784 61.576784 0.738 AA AA AA AA AA AA AA AA AA AA 1690420 1 SNP_A- rs2765249 62441479 62.441479 0.75 AA AA AA AA AA AA BB BB AA AB 1727043 1 SNP_A- rs3861943 63439667 63.439667 0.405 BB BB BB BB BB BB AB AB AA AB 1646105 1 SNP_A- rs592298 64000081 64.000081 0.25 BB BB BB BB BB BB AB AB BB AB 1654674 1 SNP_A- rs746633 64503887 64.503887 0.692 AA AA BB BB BB BB BB BB AA AB 1708628 1 SNP_A- rs1171279 65700514 65.700514 0.345 BB BB BB BB BB BB BB BB BB AB 1713897 1 SNP_A- rs1280310 66928844 66.928844 0.655 AB AB AB BB AA AB AB AB AA AB 1717648 1 SNP_A- rs1408956 67849084 67.849084 0.536 AB AB AB AA BB AB AB AB BB BB 1712508 1 SNP_A- rs1413953 70834525 70.834525 0.571 AB AB AA AA AA AA AB AB AA AB 1688631 1 SNP_A- rs1338655 73569634 73.569634 0.429 AB AB AA AA AA AA AA AA BB BB 1720162 1 SNP_A- rs10493539 74427598 74.427598 0.25 AA AA BB BB BB BB BB BB AA AB 1697494 1 SNP_A- rs277355 75002805 75.002805 0.345 BB BB BB BB BB BB AB AB BB AB 1649261 1 SNP_A- rs1250876 75905253 75.905253 0.345 AB AB AB BB AA AB AB AB BB BB 1744876 1 SNP_A- rs3928852 76926021 76.926021 0.607 AA AA AA AA AA AA AB AB AA AA 1739854 1 SNP_A- rs10493596 77438262 77.438262 0.718 AA AA AA AA AA AA AB AB AA AA 1687047 1 SNP_A- rs1248480 79071260 79.07126 0.357 BB BB AA AA AA AA AB AB BB BB 1732619 1 SNP_A- rs2127436 79792017 79.792017 0.488 AB AB AB BB AA AB BB BB AA AB 1664985 1 SNP_A- rs2389016 80511350 80.51135 0.738 AA AA AB BB AA AB AB AB AA AB 1644541 1 SNP_A- rs10518660 82094088 82.094088 0.738 AB AB AA AA AA AA AA AA AA AA 1645927 1 SNP_A- rs6598991 82697988 82.697988 0.524 BB BB AB AA AB AB AA AA AA AA 1693780 1 SNP_A- rs2268667 85505767 85.505767 0.321 BB BB AB AA AB AB BB BB BB AB 1674234 1 SNP_A- rs306322 88673430 88.67343 0.726 AA AA AA AA AA AA AB AB BB AB 1752288 1 SNP_A- rs1831298 90211840 90.21184 0.262 AB AB BB BB BB BB AA AA BB AB 1736094 1 SNP_A- rs4233429 90811924 90.811924 0.25 BB BB AB BB AB AB BB BB BB BB 1711115 1 SNP_A- rs665484 91375951 91.375951 0.512 AB AB AA AA AA AA AA AA BB BB 1714794 1 SNP_A- rs490800 92304926 92.304926 0.369 BB BB AB BB AB AB AB AB BB AB 1675488 1 SNP_A- rs6703310 93500761 93.500761 0.393 AB AB AB AA AB AB BB BB BB BB 1656572 1 SNP_A- rs223237 96276742 96.276742 0.476 BB BB BB BB BB BB AA AA AA AA 1755223 1 SNP_A- rs1911500 98291841 98.291841 0.738 AA AA AA AA AA AA AB AB AA AB 1691383 1 SNP_A- rs1838587 101983453 101.983453 0.525 AA AA AB AA AB AB AB AB AA AB 1725993 1 SNP_A- rs7521799 102944538 102.944538 0.619 BB BB AB AA AB AB AA AB BB BB 1689489 1 SNP_A- rs1576516 104210964 104.210964 0.452 AB AB AA AA AA AA BB BB AA AA 1684273 1 SNP_A- rs1919894 107240697 107.240697 0.381 BB BB BB BB BB BB AA AB BB BB 1733369 1 SNP_A- rs10494081 108114145 108.114145 0.667 AA AA AA AA AA AA AA AB BB AB 1715038 1 SNP_A- rs2026485 108978565 108.978565 0.333 BB BB AB BB AB AB BB BB BB AB 1699288 1 SNP_A- rs6682717 110527665 110.527665 0.441 AA AA AB BB AB AB AA AB AA AA 1750726 1 SNP_A- rs694180 111438255 111.438255 0.464 AB AB BB BB BB BB BB BB AA AB 1648811 1 SNP_A- rs1936061 112187978 112.187978 0.595 AA AA AA AA AA AA BB AB BB BB 1753842 1 SNP_A- rs2359417 113685242 113.685242 0.452 BB BB BB BB BB BB AA AB BB BB 1689065 1 SNP_A- rs3767824 118152606 118.152606 0.429 BB BB BB BB BB BB BB BB BB BB 1746401 1 SNP_A- rs1766803 119095860 119.09586 0.366 BB AB BB BB BB BB BB BB BB BB 1688653 1 SNP_A- rs477992 119969618 119.969618 0.286 AA AB BB BB BB AB BB BB BB AB 1708513 1 SNP_A- rs10494240 143040559 143.040559 0.321 AB AB AA AA AA AA BB AB AA AB 1701244 1 SNP_A- rs10494267 148366991 148.366991 0.321 BB BB BB BB BB BB AB AB BB BB 1706430 1 SNP_A- rs2879490 149760236 149.760236 0.381 BB BB BB BB BB AB AA AA AA AA 1680305 1 SNP_A- rs10494303 150706096 150.706096 0.571 BB BB AA AA AA AB AB AB AA AA 1723421 1 SNP_A- rs884664 151516798 151.516798 0.702 AA AA AA AA AA AB AA AA AA AA 1681003 1 SNP_A- rs10494315 154072954 154.072954 0.655 AB AB AA AA AA AB BB BB BB AB 1667931 1 SNP_A- rs919477 155585918 155.585918 0.25 BB BB BB BB BB BB BB BB BB AB 1647211 1 SNP_A- rs1149392 157241261 157.241261 0.286 BB BB BB BB BB BB BB BB BB BB 1662451 1 SNP_A- rs6683968 158923070 158.92307 0.571 AA AA BB BB BB BB AA AA AA AA 1695012 1 SNP_A- rs869513 159832184 159.832184 0.607 AA AA AA AA AA AA AA AA AA AA 1716490 1 SNP_A- rs4656422 161688624 161.688624 0.25 AB AB AA AA AB AB BB BB BB BB 1727494 1 SNP_A- rs4657482 162563307 162.563307 0.405 AB AB BB BB BB BB AB AB BB BB 1646555 1 SNP_A- rs2093658 164086850 164.08685 0.317 AA AA AA AA AB AB AA AA AA AA 1743854 1 SNP_A- rs1358948 165590931 165.590931 0.31 BB BB BB BB BB BB AB AB AA AB 1681011 1 SNP_A- rs2205848 166407951 166.407951 0.31 BB BB BB BB BB BB BB BB BB AB 1751990 1 SNP_A- rs10494487 167046739 167.046739 0.298 AB AB BB BB BB BB BB BB BB BB 1736240 1 SNP_A- rs3753538 168481215 168.481215 0.691 AA AA AA AA AA AA AA AA BB AB 1674510 1 SNP_A- rs10489280 169204277 169.204277 0.488 AA AA BB BB BB BB AA AA BB BB 1719434 1 SNP_A- rs989423 171514180 171.51418 0.75 AA AA AA AA AA AA AA AA AA AA 1753950 1 SNP_A- rs1359587 172487558 172.487558 0.691 AB AB BB BB AB AB AB AB AA AB 1722081 1 SNP_A- rs2861746 173128066 173.128066 0.452 AB AB AA AA AA AA AB AB BB BB 1694706 1 SNP_A- rs2493119 175995013 175.995013 0.333 BB BB BB BB BB BB BB BB AA AB 1733825 1 SNP_A- rs1281294 178629596 178.629596 0.714 AA AA AA AA AA AA AA AA AA AA 1671505 1 SNP_A- rs2274984 179839103 179.839103 0.524 AB AB AB BB AB AB AA AA BB BB 1694118 1 SNP_A- rs1184639 180355276 180.355276 0.357 BB BB BB BB BB BB BB BB BB AB 1644471 1 SNP_A- rs2840274 180942462 180.942462 0.429 AA AA BB BB BB BB AA AB BB BB 1711261 1 SNP_A- rs170885 181673406 181.673406 0.512 AB AB AA AA AA AA BB AB BB AB 1706912 1 SNP_A- rs10489701 182242226 182.242226 0.595 AA AA AB BB AB AB AA AA AA AA 1703470 1 SNP_A- rs10489756 182835425 182.835425 0.262 BB BB AB AA AB AB AA AA BB AB 1696277 1 SNP_A- rs726706 183604111 183.604111 0.429 AB AB BB BB BB BB BB BB AA AB 1744486 1 SNP_A- rs7543266 184360480 184.36048 0.595 AA AA AA AA AA AA AA AA AA AA 1693312 1 SNP_A- rs6665263 185050414 185.050414 0.452 BB BB BB BB BB BB BB BB AA AA 1739170 1 SNP_A- rs10494626 186275233 186.275233 0.464 AA AA BB BB BB BB BB BB BB AB 1726093 1 SNP_A- rs815160 186988747 186.988747 0.427 AB AB AA AA AA AA AA AA AA AA 1658415 1 SNP_A- rs1563191 187849406 187.849406 0.333 AB AB AB BB AB AB BB BB BB BB 1666089 1 SNP_A- rs1338034 188358939 188.358939 0.393 BB BB BB BB BB BB BB BB BB BB 1753798 1 SNP_A- rs4657868 190164348 190.164348 0.524 AB AB BB BB BB BB BB AB BB BB 1688981 1 SNP_A- rs10494707 191296870 191.29687 0.357 BB BB BB BB BB BB AA AB BB BB 1723115 1 SNP_A- rs822456 191826836 191.826836 0.439 BB BB BB BB BB BB BB BB AA AB 1651749 1 SNP_A- rs10494728 192402426 192.402426 0.25 AB AB BB BB BB BB AA AB BB BB 1642592 1 SNP_A- rs3762271 193802099 193.802099 0.6 AB AB AA AA AA AA BB BB AA AB 1658925 1 SNP_A- rs1927246 195048356 195.048356 0.702 AB AB AB BB AB AB AA AA AA AB 1687705 1 SNP_A- rs10494808 196821529 196.821529 0.548 AB AB AA AA AA AA BB BB BB AB 1725025 1 SNP_A- rs6667172 197375495 197.375495 0.5 BB BB AA AA AA AA AA AB AA AB 1665029 1 SNP_A- rs832174 197990176 197.990176 0.25 AA AA BB BB BB BB BB BB BB BB 1747494 1 SNP_A- rs7555556 199822633 199.822633 0.293 AB AB AB AB AB AB AA AB BB AB 1651207 1 SNP_A- rs10494844 200501548 200.501548 0.75 AA AA AA AA AA AA AA AB AA AA 1714962 1 SNP-A- rs10494852 201189443 201.189443 0.655 BB BB BB BB BB BB AA AB AA AA 1724123 1 SNP_A- rs311286 203999303 203.999303 0.286 AB AB BB BB BB BB BB BB BB BB 1673439 1 SNP_A- rs684431 204553812 204.553812 0.381 BB BB AA AB AA AB BB BB AB AB 1669116 1 SNP_A- rs2358452 208747444 208.747444 0.707 AA AA AA AA AA AA BB BB AA AA 1650733 1 SNP_A- rs340840 210516282 210.516282 0.393 AB AB BB BB BB BB BB BB AA AA 1651975 1 SNP_A- rs10494987 211525052 211.525052 0.691 AB AB AA AA AA AA AA AA AA AA 1683565 1 SNP_A- rs10495003 212237742 212.237742 0.417 AB AB BB AB BB AB AA AB AA AA 1750462 1 SNP_A- rs6604634 213109650 213.10965 0.524 BB BB AA AA AA AA BB BB AA AA 1683969 1 SNP_A- rs10495045 213806233 213.806233 0.714 AA AA AA AB AA AB BB BB AB AB 1731002 1 SNP_A- rs618171 215537693 215.537693 0.631 AB AB AA AA AA AA AA AA AB AB 1677675 1 SNP_A- rs10495156 217494419 217.494419 0.298 BB BB BB BB BB BB AA AA AB AB 1703136 1 SNP_A- rs1338077 218118775 218.118775 0.321 BB BB BB AB BB AB BB BB BB BB 1711849 1 SNP_A- rs4481859 219121051 219.121051 0.512 AB AB AA AA AA AA BB BB BB BB 1755399 1 SNP_A- rs10495236 221802391 221.802391 0.691 AB AB AA AB AA AB AB AB AB AB 1739524 1 SNP_A- rs710805 225430849 225.430849 0.429 AB AB BB AB BB AB AB AB AA AA 1710164 1 SNP_A- rs1998067 226545242 226.545242 0.298 BB BB AA AA AA AA BB BB BB BB 1688357 1 SNP_A- rs9286801 229119361 229.119361 0.476 AA AA AA AA AA AA AA AA AA AA 1732138 1 SNP_A- rs1892298 230387334 230.387334 0.714 AA AA AA AA AA AA AA AB AA AA 1747040 1 SNP_A- rs2463190 232711157 232.711157 0.441 AB AB BB AB BB AB AA AA AA AA 1717898 1 SNP_A- rs819639 233219640 233.21964 0.56 AB AB AA AA AA AA BB AB AA AA 1710935 1 SNP_A- rs2819774 234214896 234.214896 0.691 AA AA AA AA AA AA BB AB AB AB 1755297 1 SNP_A- rs6685861 235621137 235.621137 0.357 BB BB AA AA AA AA AA AB AA AA 1677233 1 SNP_A- rs732160 236262770 236.26277 0.298 BB BB BB BB BB BB BB BB AB AB 1679485 1 SNP_A- rs1039529 238918670 238.91867 0.619 AA AA AA AA AA AA AA AA AA AA 1679759 1 SNP_A- rs879725 240732087 240.732087 0.464 BB AB AA AA AA AA AA AB AB AB 1664943 1 SNP_A- rs1093961 241902498 241.902498 0.415 BB AB AB BB AB AB AA AB AB AB 1724627 1 SNP_A- rs3844080 243632874 243.632874 0.655 BB BB AA AA AA AA BB AB AA AA 1672603 2 SNP_A- rs10519439 108913 0.108913 0.274 BB BB BB BB BB BB AB AB BB BB 1753456 2 SNP_A- rs6759198 2342478 2.342478 0.56 AA AA BB BB BB BB AB AB BB BB 1746820 2 SNP_A- rs2119075 4395806 4.395806 0.607 AA AA AA AB AB AB AB AB AB AB 1697325 2 SNP_A- rs963964 5206872 5.206872 0.321 BB BB BB BB BB BB BB BB AB AB 1740868 2 SNP_A- rs1429220 5881639 5.881639 0.369 BB BB BB BB BB BB BB BB BB BB 1677893 2 SNP_A- rs6727796 7605796 7.605796 0.512 BB BB AA AA AA AA BB BB AB AB 1650909 2 SNP_A- rs9287698 8437894 8.437894 0.281 BB BB BB BB BB BB AB AB AA AA 1663651 2 SNP_A- rs2271333 9323848 9.323848 0.5 AB AB AA AA AA AA BB BB BB BB 1647101 2 SNP_A- rs2241113 10226344 10.226344 0.31 BB BB AB AB AB AB BB AB BB BB 1717786 2 SNP_A- rs1686426 10899146 10.899146 0.5 BB BB AB AB AB AB BB BB BB BB 1706150 2 SNP_A- rs4669806 12151350 12.15135 0.619 BB BB AA AA AA AA AA AA AB AB 1676173 2 SNP_A- rs625842 12779571 12.779571 0.583 BB BB AA AA AA AA BB BB AB AB 1664687 2 SNP_A- rs7568703 15041402 15.041402 0.571 AA AA AB AB AB AB BB BB AA AA 1696327 2 SNP_A- rs4668968 15835884 15.835884 0.369 BB BB BB BB BB BB BB BB BB BB 1683239 2 SNP_A- rs9306902 16971747 16.971747 0.631 AA AA AA AA AA AA AA AA AA AA 1677981 2 SNP_A- rs10495699 19918971 19.918971 0.56 AB AB AB AB AA AB AA AB AB AB 1714454 2 SNP_A- rs10495705 20662972 20.662972 0.564 AA AA AA AA AA AA BB AB AA AA 1668860 2 SNP_A- rs7594267 23344557 23.344557 0.571 AA AA BB BB BB BB BB BB AA AA 1693698 2 SNP_A- rs1275963 26804398 26.804398 0.643 AB AB AA AA AA AA AA AA AA AA 1751070 2 SNP_A- rs2014701 30210694 30.210694 0.631 AA AA AA AA AA AA BB AB BB BB 1684619 2 SNP_A- rs10490360 32207919 32.207919 0.441 BB BB AA AA AA AA BB BB AB AB 1648557 2 SNP_A- rs219145 33123165 33.123165 0.634 AA AA AA AA AA AA AA AA BB BB 1671421 2 SNP_A- rs10495796 34029149 34.029149 0.274 BB BB BB BB BB BB BB BB BB BB 1696185 2 SNP_A- rs2049638 34866446 34.866446 0.738 AA AA AA AA AA AA AA AB AB AB 1642658 2 SNP_A- rs1401242 35923474 35.923474 0.417 AB AB AB AB AA AB AA AB AB AB 1696029 2 SNP_A- rs2161905 36427824 36.427824 0.286 BB BB BB BB BB BB AA AB BB BB 1748242 2 SNP_A- rs975315 38407251 38.407251 0.631 AB AB AA AA AA AA AA AB AA AA 1660238 2 SNP_A- rs9309043 39939220 39.93922 0.643 BB BB AB AB BB AB BB BB BB BB 1719460 2 SNP_A- rs2059338 41186539 41.186539 0.714 AB AB AB AB BB AB AA AB BB BB 1714203 2 SNP_A- rs10495900 43528810 43.52881 0.369 AB AB BB BB BB BB BB BB BB BB 1690204 2 SNP_A- rs4276071 44121457 44.121457 0.417 BB BB AA AA AA AA AB AB BB BB 1740164 2 SNP_A- rs6708061 44721790 44.72179 0.357 BB BB BB BB BB BB AA AA AB AB 1685265 2 SNP_A- rs6737073 45442330 45.44233 0.286 BB BB BB BB BB BB AA AA AB AB 1680505 2 SNP_A- rs935661 45967008 45.967008 0.56 AB AB AB AA AB AB AB AB AB AB 1680749 2 SNP_A- rs7589621 46494033 46.494033 0.738 AB AB AA AA AA AA AA AA AA AA 1721531 2 SNP_A- rs6544955 47235732 47.235732 0.643 AA AA AA AA AA AA AA AA AA AA 1734011 2 SNP_A- rs10495972 49412546 49.412546 0.298 AB AB BB BB BB BB BB BB BB BB 1699364 2 SNP_A- rs10495987 50064931 50.064931 0.286 AB AB AA AA AA AA BB BB BB BB 1696353 2 SNP_A- rs10490176 50979585 50.979585 0.738 AB AB AA AA AA AA AA AA AA AA 1646009 2 SNP_A- rs1160297 53148971 53.148971 0.321 AB AB BB BB BB BB BB BB AB AB 1676509 2 SNP_A- rs843622 54460133 54.460133 0.524 BB BB AA AA AA AA BB BB AA AA 1742892 2 SNP_A- rs5008666 59390639 59.390639 0.65 AA AA BB BB BB BB AA AA AA AB 1704006 2 SNP_A- rs1517401 63452358 63.452358 0.738 AA AA BB BB BB BB AB AB AA AA 1673301 2 SNP_A- rs2581047 64219699 64.219699 0.286 BB BB BB BB BB BB BB BB BB BB 1711471 2 SNP_A- rs2971828 66466238 66.466238 0.667 BB BB AA AA AA AA AA AA AA AA 1652253 2 SNP_A- rs9309400 67746695 67.746695 0.738 AB AB AA AA AA AA BB BB AA AA 1722279 2 SNP_A- rs10496165 68531740 68.53174 0.333 BB BB BB BB BB BB BB BB AA AB 1656216 2 SNP_A- rs2312209 69633766 69.633766 0.524 BB BB AA AA AB AB BB BB BB BB 1703772 2 SNP_A- rs10489986 70719802 70.719802 0.75 AA AA AA AA AA AA AB AB AA AA 1753748 2 SNP_A- rs6724782 73591645 73.591645 0.286 AA AB BB BB BB BB AB AB BB BB 1723065 2 SNP_A- rs730148 76269470 76.26947 0.408 BB BB AA AA AB AB AA AA BB BB 1665193 2 SNP_A- rs1446707 77141310 77.14131 0.429 BB BB AA AA AA AA AA AA BB BB 1718918 2 SNP_A- rs4852483 79513076 79.513076 0.75 AA AB BB BB BB BB AA AA AA AA 1749208 2 SNP_A- rs216616 80715814 80.715814 0.298 BB BB AA AA AA AA BB BB BB BB 1745171 2 SNP_A- rs9309572 81381197 81.381197 0.321 BB BB BB BB BB AB AA AA BB AB 1750234 2 SNP_A- rs7577293 85846940 85.84694 0.274 AA AB BB BB BB BB BB BB AA AB 1728812 2 SNP_A- rs9308826 99738211 99.738211 0.738 AA AA AA AA AA AA AA AA AA AA 1676217 2 SNP_A- rs9308849 101983905 101.983905 0.714 AA AA AA AA AA AA AA AB BB AB 1655416 2 SNP_A- rs956966 103046093 103.046093 0.512 AA AA BB BB BB AB BB BB BB BB 1655538 2 SNP_A- rs1869070 106074094 106.074094 0.714 AA AA AA AA AA AA AB AB AB AB 1690274 2 SNP_A- rs1398132 106705516 106.705516 0.607 AB AB AA AA AA AA BB BB AB AB 1742362 2 SNP_A- rs826690 108705477 108.705477 0.429 BB BB AA AA AB AB AB AB AB AB 1654768 2 SNP_A- rs1469529 109207139 109.207139 0.583 AB AB AA AA AB AB AB AB AA AA 1709888 2 SNP_A- rs3961919 112959552 112.959552 0.298 BB BB AA AA AB AB AB AB AB AB 1671489 2 SNP_A- rs2166965 114191141 114.191141 0.679 AA AA AA AA AB AB AA AA AA AA 1720080 2 SNP_A- rs1346762 114988791 114.988791 0.72 AA AA AA AA AA AA AA AA AA AA 1712138 2 SNP_A- rs9284719 118395025 118.395025 0.595 AB AB AA AA AA AA AA AA AA AA 1752188 2 SNP_A- rs1370380 120731125 120.731125 0.286 AB AB AA AA AA AA BB BB AB AB 1685413 2 SNP_A- rs4848174 122659698 122.659698 0.655 AA AA AA AA AB AB AB AB AA AA 1721631 2 SNP_A- rs1215318 125809045 125.809045 0.536 AA AA BB BB BB BB BB BB AB AB 1707304 2 SNP_A- rs548032 127461866 127.461866 0.631 AA AA AA AA AA AA AA AA AB AB 1673583 2 SNP_A- rs2124432 128900396 128.900396 0.61 AB AB AA AA AA AA AA AA AA AA 1671177 2 SNP_A- rs10496731 135431360 135.43136 0.488 BB BB AB BB AB AB AB AB BB BB 1676259 2 SNP_A- rs10496750 137176540 137.17654 0.667 AB AB AB AA AB AB AA AA AB AB 1689435 2 SNP_A- rs10490739 137712397 137.712397 0.287 BB BB BB BB BB BB BB BB BB BB 1695208 2 SNP_A- rs3884566 139082237 139.082237 0.357 AB AB BB BB BB BB AB AB AB AB 1651715 2 SNP_A- rs3922799 139592638 139.592638 0.476 BB BB BB BB BB BB BB BB AA AA 1665733 2 SNP_A- rs838042 140153918 140.153918 0.262 AA AA AB AA AB AB BB BB BB BB 1713885 2 SNP_A- rs1518441 140908218 140.908218 0.286 AA AA BB BB BB BB AB AB AB AB 1663529 2 SNP_A- rs10496859 141502410 141.50241 0.536 BB BB AA AA AA AA AA AA BB BB 1643152 2 SNP_A- rs355562 142245134 142.245134 0.321 BB BB BB BB BB BB BB BB BB BB 1689866 2 SNP_A- rs7560400 143121832 143.121832 0.75 AA AA AB BB AB AB AB AB AA AA 1688373 2 SNP_A- rs1437717 146329325 146.329325 0.571 AB AB AB BB AB AB AB AB AA AA 1725903 2 SNP_A- rs1528842 148291308 148.291308 0.75 AB AB AB BB AB AB AA AA AA AA 1729119 2 SNP_A- rs6734792 151450390 151.45039 0.738 AA AA AA AA AA AA BB BB AA AA 1716616 2 SNP_A- rs9287956 151979514 151.979514 0.464 BB BB AA AA AA AA AB AB AB AB 1645341 2 SNP_A- rs1370502 153235438 153.235438 0.667 BB BB BB BB BB BB AA AA AB AB 1711079 2 SNP_A- rs10497129 153977886 153.977886 0.31 AB AB AA AA AA AA BB BB BB BB 1751360 2 SNP_A- rs1469155 155088509 155.088509 0.726 AA AA AA AA AA AA BB BB AB AB 1682179 2 SNP_A- rs6750583 159423695 159.423695 0.738 BB BB AB BB AA AB AB AB AB AB 1729675 2 SNP_A- rs997163 161593412 161.593412 0.366 AA AA BB BB BB BB AA AA BB BB 1710753 2 SNP_A- rs1227921 162517707 162.517707 0.512 BB BB AB BB AA AB AB AB BB BB 1657420 2 SNP_A- rs1446471 164812395 164.812395 0.345 BB BB AB AA BB AB AA AA BB BB 1681353 2 SNP_A- rs10497261 166152395 166.152395 0.702 AB AB AA AA AA AA AB AB BB AB 1755647 2 SNP_A- rs9287874 167411538 167.411538 0.738 AA AA AA AA AA AA AB AB BB AB 1656096 2 SNP_A- rs2278785 168822282 168.822282 0.381 BB BB BB BB BB BB AA AA BB BB 1673653 2 SNP_A- rs830995 169955143 169.955143 0.702 AA AA AA AA AA AA AB AB AA AB 1702574 2 SNP_A- rs961313 170759024 170.759024 0.274 BB BB BB BB BB BB AB AB BB BB 1645337 2 SNP_A- rs731693 171622741 171.622741 0.667 AA AA AA AA AA AA AB AB BB BB 1687817 2 SNP_A- rs4095835 172330518 172.330518 0.429 BB BB BB BB BB BB BB BB BB AB 1749036 2 SNP_A- rs7575189 173840675 173.840675 0.512 BB BB AB BB AA AB AB AB BB BB 1683223 2 SNP_A- rs2119137 174843221 174.843221 0.333 AB AB AB AA BB AB AB AB BB BB 1743510 2 SNP_A- rs1993385 175563974 175.563974 0.476 AA AA AB BB AA AB AB AB AA AB 1673703 2 SNP_A- rs9287989 176543248 176.543248 0.643 BB BB AA AA AA AA AA AA AA AB 1730586 2 SNP_A- rs6722762 177140660 177.14066 0.345 AB AB BB BB BB BB AB AB AA AB 1676261 2 SNP_A- rs10497467 177733918 177.733918 0.75 AA AA AB BB AA AB AB AB AA AA 1668972 2 SNP_A- rs2008999 179796838 179.796838 0.643 AA AA BB BB BB BB AA AA AA AA 1643400 2 SNP_A- rs259845 180560120 180.56012 0.75 AA AA AA AA AA AA BB BB BB BB 1721647 2 SNP_A- rs9288052 181299542 181.299542 0.488 BB BB BB BB BB BB BB BB BB BB 1643999 2 SNP_A- rs288332 183450856 183.450856 0.262 BB BB AA AA AA AA AA AA AA AB 1668465 2 SNP_A- rs1454042 184407382 184.407382 0.357 BB BB BB BB BB BB AB AB AA AB 1723211 2 SNP_A- rs10490389 186428458 186.428458 0.702 AA AA AA AA AA AA AA AA AA AA 1668055 2 SNP_A- rs2044683 187026818 187.026818 0.366 BB BB BB BB BB BB AB AB BB BB 1678177 2 SNP_A- rs840611 188023952 188.023952 0.583 BB BB BB BB BB BB BB BB AA AA 1728072 2 SNP_A- rs10497725 192818722 192.818722 0.667 AA AA BB BB BB BB AA AA AA AB 1750900 2 SNP_A- rs10497744 194316402 194.316402 0.31 BB BB BB BB BB BB BB BB BB AB 1642958 2 SNP_A- rs1350208 198911771 198.911771 0.571 AB AB AB BB AB AB AA AA AA AB 1669242 2 SNP_A- rs10497821 199463403 199.463403 0.31 BB BB BB BB BB BB BB BB AA AB 1673517 2 SNP_A- rs1376877 204097596 204.097596 0.607 AA AA BB BB BB BB BB BB BB AB 1645863 2 SNP_A- rs6707500 204941128 204.941128 0.667 AB AB AA AA AA AA AA AA AA AA 1650883 2 SNP_A- rs10490293 206049378 206.049378 0.274 BB BB AA AA AA AA BB BB BB BB 1757786 2 SNP_A- rs10490474 207934338 207.934338 0.571 BB BB AB AA AB AB AB AB AA AA 1752790 2 SNP_A- rs10497888 208586741 208.586741 0.679 AA AA BB BB BB BB AA AA AA AA 1642246 2 SNP_A- rs1607181 209364109 209.364109 0.655 AA AB AA AA AA AA AB AB BB AB 1644145 2 SNP_A- rs1816532 212093746 212.093746 0.75 AA AA BB BB BB BB AA AA AA AA 1669816 2 SNP_A- rs1402769 212906949 212.906949 0.274 BB BB BB BB BB BB BB BB BB AB 1661335 2 SNP_A- rs10497986 213664725 213.664725 0.702 AA AA BB BB BB BB AB AB AA AA 1720206 2 SNP_A- rs9283527 214674151 214.674151 0.417 AA AA AB BB AB AB AA AA AA AA 1701518 2 SNP_A- rs2166459 215505298 215.505298 0.31 AA AB BB BB BB BB AA AA BB AB 1692929 2 SNP_A- rs1250225 216151895 216.151895 0.744 BB AB AA AA AA AA AA AA AA AA 1755667 2 SNP_A- rs1110998 217169458 217.169458 0.429 BB BB AB AA AB AB BB BB BB BB 1705890 2 SNP_A- rs6719545 218277340 218.27734 0.286 BB AB BB BB BB BB BB BB AA AB 1743410 2 SNP_A- rs1344645 219363524 219.363524 0.405 AA AB BB BB BB BB AA AA BB BB 1728154 2 SNP_A- rs715345 220649461 220.649461 0.274 AA AA AB AB AB AB BB BB BB BB 1726837 2 SNP_A- rs1356399 221348562 221.348562 0.702 AA AA AB AB AB AB BB AB AA AA 1721280 2 SNP_A- rs1430234 222049201 222.049201 0.274 BB BB BB BB BB BB BB BB BB AB 1655920 2 SNP_A- rs4673013 222802583 222.802583 0.619 BB AB AB AB AB AB AA AA AA AB 1711521 2 SNP_A- rs1961637 223730613 223.730613 0.452 AA AB AB AB AB AB BB BB BB BB 1695224 2 SNP_A- rs10498158 224777152 224.777152 0.417 AA AB BB BB BB BB BB BB BB BB 1713318 2 SNP_A- rs10498171 225622115 225.622115 0.524 BB AB AA AA AA AA BB AB BB AB 1702406 2 SNP_A- rs1835533 226193946 226.193946 0.738 AA AA AA AA AA AA AA AA AA AA 1643000 2 SNP_A- rs1522804 226814661 226.814661 0.548 AA AB AB AB AB AB BB BB BB AB 1739924 2 SNP_A- rs1950134 227888457 227.888457 0.714 AA AA AA AA AA AA AA AA AA AA 1683533 2 SNP_A- rs1524023 228615089 228.615089 0.75 AA AA AA AA AA AA AA AA AA AA 1663421 2 SNP_A- rs6759815 229797926 229.797926 0.571 BB BB AA AA AA AA BB BB BB AB 1707748 2 SNP_A- rs4973304 230936568 230.936568 0.298 BB BB BB BB BB BB AA AA AA AB 1661533 2 SNP_A- rs10498257 231818543 231.818543 0.702 BB AB AB AB AB AB BB AB AA AA 1727506 2 SNP_A- rs3791711 233258388 233.258388 0.524 BB AB BB BB BB BB BB AB AA AB 1728658 2 SNP_A- rs1880747 235240125 235.240125 0.512 BB AB AA AA AA AA BB AB AA AA 1747120 2 SNP_A- rs103718 239180488 239.180488 0.441 AA AA AA AA AA AA AB AB AA AA 1738177 3 SNP_A- rs1516342 147906 0.147906 0.262 AB AB BB BB BB BB AA AB BB BB 1675236 3 SNP_A- rs10510204 981912 0.981912 0.405 AA AA AA AA AA AA AA AB AA AA 1754907 3 SNP_A- rs1720194 2366613 2.366613 0.631 AA AA AA AA AA AA AA AA AA AA 1726483 3 SNP_A- rs1508734 3538991 3.538991 0.607 AB AB AB BB BB AB BB AB AB AB 1717220 3 SNP_A- rs4684484 5437541 5.437541 0.441 BB BB AA AA AA AA AA AA AA AA 1668475 3 SNP_A- rs9311817 6172932 6.172932 0.72 AB AB AA AA AA AA AA AA AB AB 1646075 3 SNP_A- rs1450097 7520521 7.520521 0.293 BB BB BB BB BB BB AB AB AB AB 1658187 3 SNP_A- rs486012 9016299 9.016299 0.56 AA AA AA AA AA AA AB AB AA AB 1718736 3 SNP_A- rs2160871 10421826 10.421826 0.75 AB AB AB BB BB AB AA AA AA AB 1679373 3 SNP_A- rs6792718 11409380 11.40938 0.429 AB AB AA AA AA AA AA AA AA AA 1649097 3 SNP_A- rs172429 14880517 14.880517 0.25 BB BB BB BB BB BB BB BB BB BB 1713577 3 SNP_A- rs1983085 15504547 15.504547 0.56 AA AA BB BB BB BB AB AB AA AA 1719120 3 SNP_A- rs2733528 17211259 17.211259 0.571 AB AB AA AA AA AA AB AB BB BB 1701284 3 SNP_A- rs336615 18605807 18.605807 0.619 AA AA AB BB BB AB AB AB BB AB 1653347 3 SNP_A- rs2053506 19350795 19.350795 0.595 AA AA AA AA AA AA AA AA BB BB 1753110 3 SNP_A- rs6770717 20406548 20.406548 0.726 AB AB AA AA AA AA AA AA AA AA 1649119 3 SNP_A- rs365392 21465872 21.465872 0.583 BB BB AB BB BB AB AB AB BB AB 1685927 3 SNP_A- rs3732395 23209622 23.209622 0.378 AA AB BB BB BB BB BB BB BB AB 1738848 3 SNP_A- rs10510568 25577736 25.577736 0.679 BB AB AA AA AA AA AB AB AA AB 1730534 3 SNP_A- rs9284859 26883268 26.883268 0.655 BB BB AB AB BB AB AB AB BB AB 1725077 3 SNP_A- rs7639905 27951868 27.951868 0.429 BB AB BB BB BB BB BB BB AA AA 1647333 3 SNP_A- rs9310901 29477393 29.477393 0.274 BB AB AA AA AA AA AB AB BB AB 1744932 3 SNP_A- rs795347 30720945 30.720945 0.369 AA AB AB AB AB AB BB BB BB BB 1741570 3 SNP_A- rs347163 32435579 32.435579 0.393 BB BB BB BB BB BB BB BB AB AB 1747050 3 SNP_A- rs1376015 35274750 35.27475 0.595 BB AB AB AB AB AB AB AB BB BB 1735191 3 SNP_A- rs10510667 35834447 35.834447 0.476 BB BB AA AA AA AA BB BB AB AB 1717686 3 SNP_A- rs10510695 37621200 37.6212 0.738 AA AA AA AA AA AA AA AA AA AA 1643995 3 SNP_A- rs2220345 41411812 41.411812 0.429 AA AB AA AA AA AA AB AB AB AB 1649705 3 SNP_A- rs531888 43047989 43.047989 0.476 BB AB AB AB AB AB AB AB AA AA 1699750 3 SNP_A- rs2742393 45732421 45.732421 0.417 AA AA AB AB AB AB BB BB AA AA 1722715 3 SNP_A- rs7620394 55206368 55.206368 0.345 BB BB AA AA AA AA BB BB AB AB 1694360 3 SNP_A- rs6445844 57028961 57.028961 0.726 BB BB AB AB AB AB AB AB AA AA 1643909 3 SNP_A- rs10510803 59329572 59.329572 0.488 BB BB AB AB AB AB AB AB AB AB 1652229 3 SNP_A- rs3843360 60016727 60.016727 0.393 BB BB AB AB AB AB AB AB AA AA 1669748 3 SNP_A- rs1996520 61592725 61.592725 0.488 BB BB BB BB BB BB AB AB AB AB 1678019 3 SNP_A- rs7650561 62466087 62.466087 0.643 AA AA AB AB AB AB AA AA AB AB 1665709 3 SNP_A- rs10510929 64709076 64.709076 0.583 BB BB BB BB BB BB AA AA BB AB 1684953 3 SNP_A- rs725160 66943022 66.943022 0.464 AA AA BB BB BB BB AB AB BB BB 1688393 3 SNP_A- rs4145917 68099517 68.099517 0.679 AB AB AA AA AA AA AA AA AA AA 1678015 3 SNP_A- rs2872939 69802192 69.802192 0.405 AB AB AA AA AA AA AB AB BB BB 1707438 3 SNP_A- rs10510996 70545357 70.545357 0.75 AA AA AA AB AB AB AA AA AA AB 1663707 3 SNP_A- rs830644 71748249 71.748249 0.5 AB AB BB BB BB BB BB BB AA AB 1650625 3 SNP_A- rs4677226 73154304 73.154304 0.613 BB BB BB BB BB BB AB AB AA AA 1713028 3 SNP_A- rs1107768 73959415 73.959415 0.726 AB AB AA AA AA AA BB AB AA AA 1685633 3 SNP_A- rs10511039 76184447 76.184447 0.583 AA AA BB BB BB BB AA AA BB BB 1722733 3 SNP_A- rs251552 76852596 76.852596 0.539 AB AB AA AA AA AA AA AB BB AB 1648479 3 SNP_A- rs9309840 80029943 80.029943 0.588 BB BB BB BB BB BB BB AB BB BB 1642486 3 SNP_A- rs2639611 81623522 81.623522 0.274 BB BB BB BB BB BB BB BB AA AA 1685115 3 SNP_A- rs9309888 82418655 82.418655 0.262 BB AB BB BB BB BB BB BB AA AB 1642250 3 SNP_A- rs10511085 85614577 85.614577 0.619 BB AB BB AB BB AB AA AA AA AB 1732971 3 SNP_A- rs1509783 87634505 87.634505 0.476 BB BB BB BB BB BB AA AB BB BB 1731608 3 SNP_A- rs9310061 88146455 88.146455 0.631 AA AA BB BB BB AB AA AA AA AB 1721601 3 SNP_A- rs724972 89664098 89.664098 0.607 AA AB AA AA AA AA AA AA AA AA 1715294 3 SNP_A- rs10511152 96638708 96.638708 0.381 BB BB AA AA AA AB AA AB BB AB 1648583 3 SNP_A- rs3856571 99031739 99.031739 0.298 BB AB BB BB BB BB BB BB BB BB 1701406 3 SNP_A- rs10511169 100116062 100.116062 0.691 AA AA AA AA AA AA AA AB BB AB 1643841 3 SNP_A- rs2700633 100643241 100.643241 0.643 AA AB AA AA AA AA AA AA BB AB 1697988 3 SNP_A- rs10511183 102046105 102.046105 0.738 AA AA AA AA AA AA AA AA AA AA 1740468 3 SNP_A- rs974059 103277828 103.277828 0.25 BB BB BB BB BB BB AA AB BB AB 1746982 3 SNP_A- rs1391423 103923668 103.923668 0.732 AA AA AA AA AA AA AA AA AA AA 1687227 3 SNP_A- rs10511221 105099054 105.099054 0.726 BB AB AA AA AB AB AA AA AA AB 1677819 3 SNP_A- rs6783422 106031580 106.03158 0.393 AA AA AA AA AA AA BB BB AA AA 1663937 3 SNP_A- rs10511243 106653352 106.653352 0.667 AA AA AA AA AA AA BB AB AA AB 1674588 3 SNP_A- rs2222039 108202685 108.202685 0.691 BB AB AA AA AA AA AA AA AA AA 1652015 3 SNP_A- rs1525873 111232702 111.232702 0.702 BB AB BB BB BB BB AA AA AA AB 1722407 3 SNP_A- rs1512514 111766406 111.766406 0.488 BB AB AA AA AA AA AA AA AA AA 1674512 3 SNP_A- rs1797626 114308943 114.308943 0.702 AA AA BB BB AB AB AA AA AA AA 1747616 3 SNP_A- rs1553209 116705663 116.705663 0.476 BB AB AA AA AA AA BB BB BB BB 1668954 3 SNP_A- rs7621196 117804184 117.804184 0.321 BB BB AA AA AA AA BB AB AA AB 1674292 3 SNP_A- rs1218621 118459636 118.459636 0.31 AA AA BB BB BB BB BB BB BB BB 1643903 3 SNP_A- rs950649 121567065 121.567065 0.691 BB AB AA AA AA AA AA AA AA AA 1728638 3 SNP_A- rs2126140 122627958 122.627958 0.5 BB AB AB AA AB AB AB AB AA AA 1730195 3 SNP_A- rs10511409 123610479 123.610479 0.738 BB AB AA AA AA AA AB AB BB AB 1741126 3 SNP_A- rs1373606 125496637 125.496637 0.342 AA AA BB BB BB BB AA AA AA AA 1739520 3 SNP_A- rs1374804 127391196 127.391196 0.524 AA AB AB AA AB AB BB BB AA AA 1727336 3 SNP_A- rs2718880 132343455 132.343455 0.75 AA AA AA AA AA AA AB AB AA AA 1683659 3 SNP_A- rs1553975 132999274 132.999274 0.369 BB BB AB AA AB AB AB AB BB BB 1747192 3 SNP_A- rs2310229 133541978 133.541978 0.691 AA AA AA AA AA AA AA AA AA AA 1744702 3 SNP_A- rs711923 136539253 136.539253 0.744 AA AA AA AA AA AA AA AA AA AA 1730051 3 SNP_A- rs838623 144671624 144.671624 0.619 AA AA AA AA AA AA AA AA BB AB 1654278 3 SNP_A- rs4610179 146387799 146.387799 0.726 AB AB AA AA AA AA AA AA AA AA 1730514 3 SNP_A- rs4592991 147111601 147.111601 0.393 BB BB BB BB BB BB AA AA AB AB 1700733 3 SNP_A- rs7645488 149410366 149.410366 0.31 AB AB BB BB BB BB BB BB AB AB 1744174 3 SNP_A- rs2130319 150976344 150.976344 0.333 AB AB BB BB BB BB AA AA AB AB 1718574 3 SNP_A- rs7648424 151906089 151.906089 0.488 AB AB BB BB BB BB AA AB AB AB 1718772 3 SNP_A- rs10513399 152600180 152.60018 0.488 BB BB BB BB BB BB BB BB AB AB 1663723 3 SNP_A- rs2418925 155234610 155.23461 0.524 AA AA AB AB AA AB BB BB BB BB 1746211 3 SNP_A- rs6772323 157710345 157.710345 0.667 AA AA AA AA AA AA AA AA AA AA 1658251 3 SNP_A- rs4679851 160261035 160.261035 0.274 AB AB BB BB BB BB BB AB AB AB 1736960 3 SNP_A- rs10513549 161237209 161.237209 0.25 BB BB AB AB AA AB BB AB AB AB 1716368 3 SNP_A- rs336583 162564683 162.564683 0.417 AA AA BB BB BB BB BB BB BB BB 1726685 3 SNP_A- rs7635791 163720371 163.720371 0.655 AA AA AB AB AA AB BB AB AB AB 1721879 3 SNP_A- rs9290201 164397051 164.397051 0.31 AB AB AB AB AA AB BB AB AB AB 1745785 3 SNP_A- rs4352381 165179142 165.179142 0.369 BB BB AA AA AA AA AA AB AB AB 1697475 3 SNP_A- rs2643191 165861395 165.861395 0.524 AA AA AA AA AA AA BB BB BB BB 1748578 3 SNP_A- rs1371900 167443656 167.443656 0.286 AB AB AB AB AA AB BB BB AB AB 1687865 3 SNP_A- rs1877269 170109722 170.109722 0.548 AB AB AB AB AA AB AB AB AB AB 1680949 3 SNP_A- rs8192675 172207585 172.207585 0.732 AB AB AB AB BB AB AA AA AA AA 1731022 3 SNP_A- rs7627220 173288405 173.288405 0.441 AB AB AA AA AA AA BB BB AB AB 1656780 3 SNP_A- rs792354 174456847 174.456847 0.357 BB BB AA AA AA AA BB BB BB BB 1720350 3 SNP_A- rs1377828 177727744 177.727744 0.286 AA AA BB BB BB BB BB BB BB BB 1662989 3 SNP_A- rs2160836 179192927 179.192927 0.662 AB AB BB BB BB BB AA AA AA AA 1651103 3 SNP_A- rs6762743 180494702 180.494702 0.667 AA AA AA AA AA AA AA AA AB AB 1699226 3 SNP_A- rs262958 184975690 184.97569 0.583 AB AB AA AA AA AA BB BB AB AB 1655724 3 SNP_A- rs10513799 186032241 186.032241 0.75 AB AB AA AA AA AA AB AB AB AB 1726281 3 SNP_A- rs1962838 189742951 189.742951 0.405 AB AB AB AB AB AB AA AA AB AB 1649485 3 SNP_A- rs2378464 190305279 190.305279 0.262 AB AB BB BB BB BB AB AB BB BB 1756920 3 SNP_A- rs3773928 191066407 191.066407 0.405 BB BB AB AB AB AB BB BB AB AB 1734403 3 SNP_A- rs1405036 192749559 192.749559 0.262 AB AB AB AB AB AB BB BB BB BB 1720858 3 SNP_A- rs1403033 193538911 193.538911 0.441 AB AB AA AA AA AA AB AB AB AB 1706600 3 SNP_A- rs587612 195020261 195.020261 0.369 AA AA BB BB BB BB AA AA BB BB 1643612 4 SNP_A- rs1059159 5647306 5.647306 0.683 AB AB AA AA AA AA AA AA BB BB 1669560 4 SNP_A- rs10489076 9947117 9.947117 0.691 AB AB AA AA AA AA AB AB AA AB 1743690 4 SNP_A- rs959233 10578428 10.578428 0.452 AA AA AA AA AA AA AB AB BB BB 1736300 4 SNP_A- rs10516254 12310930 12.31093 0.714 AA AA AB AB AB AB BB BB BB AB 1750658 4 SNP_A- rs10489092 13327021 13.327021 0.286 AB AB AA AA AA AA AB AB AA AB 1712820 4 SNP_A- rs10488982 14088975 14.088975 0.5 AB AB AB AA AB AB BB BB BB BB 1709160 4 SNP_A- rs1496747 16275503 16.275503 0.476 BB BB AB BB AB AB AB AB BB BB 1748456 4 SNP_A- rs10516339 19549340 19.54934 0.725 AA AB AA AA AA AA AA AA AA AA 1674656 4 SNP_A- rs6834573 20123113 20.123113 0.298 BB BB BB BB BB BB BB BB BB BB 1659171 4 SNP_A- rs10516397 21369936 21.369936 0.405 BB BB AB BB AB AB BB BB AA AA 1687559 4 SNP_A- rs2036713 22984189 22.984189 0.357 BB AB AB BB AB AB BB BB BB AB 1695570 4 SNP_A- rs1527354 24561836 24.561836 0.655 BB AB BB BB BB BB AA AA BB BB 1649429 4 SNP_A- rs7697266 25453418 25.453418 0.393 BB BB BB BB BB BB BB BB BB BB 1710973 4 SNP_A- rs9291495 27032051 27.032051 0.75 AA AA AA AA AA AA AA AA AA AB 1748352 4 SNP_A- rs1397438 28093488 28.093488 0.463 BB BB BB BB BB BB BB BB AA AA 1737486 4 SNP_A- rs939573 28670407 28.670407 0.75 AA AA AA AA AA AA AA AA AA AA 1660740 4 SNP_A- rs1441691 29221732 29.221732 0.274 BB BB AB BB AB AB BB BB BB AB 1659069 4 SNP_A- rs2571468 29891942 29.891942 0.667 AA AA AB AA AB AB AB AB AA AA 1731582 4 SNP_A- rs412253 31119019 31.119019 0.72 AA AA AB BB AB AB AB AB AA AA 1666099 4 SNP_A- rs10517232 31725815 31.725815 0.321 BB BB AA AA AA AA BB BB AA AA 1659419 4 SNP_A- rs2588544 36822899 36.822899 0.281 AA AB BB BB BB BB BB BB BB BB 1743944 4 SNP_A- rs7693744 42094241 42.094241 0.488 AA AA AB AA AA AB AA AB AA AA 1650541 4 SNP_A- rs10517054 42743857 42.743857 0.726 AA AA AA AA AA AA AA AA AA AA 1651577 4 SNP_A- rs10517094 44153139 44.153139 0.31 BB BB BB BB BB BB BB BB BB BB 1708293 4 SNP_A- rs10517121 44712712 44.712712 0.583 AA AA AB AA AA AB AA AB BB AB 1672145 4 SNP_A- rs1552419 45366813 45.366813 0.583 AA AB AA AA AA AA BB AB AA AA 1742914 4 SNP_A- rs279842 46181884 46.181884 0.439 BB BB BB BB BB BB BB AB AA AB 1741538 4 SNP_A- rs3934674 46854066 46.854066 0.305 BB BB BB BB BB BB BB BB BB BB 1726797 4 SNP_A- rs6447614 47908885 47.908885 0.549 AA AB AA AA AA AB AA AA BB AB 1734487 4 SNP_A- rs6850277 54268853 54.268853 0.667 AB AB AA AA AA AA AA AB AA AA 1659623 4 SNP_A- rs2726610 55528245 55.528245 0.548 BB BB BB BB BB BB BB AB AA AB 1724073 4 SNP_A- rs4580704 56167635 56.167635 0.643 AB AB AA AA AA AA AA AA AA AB 1643184 4 SNP_A- rs10517400 58338522 58.338522 0.679 AB AB BB BB AB AB AB AB BB BB 1647321 4 SNP_A- rs10517453 60065841 60.065841 0.679 AA AA BB BB BB BB AB AB BB BB 1685901 4 SNP_A- rs2129274 61712878 61.712878 0.613 BB BB BB BB BB BB BB BB AA AA 1660836 4 SNP_A- rs2345043 62476674 62.476674 0.619 AA AA AA AA AA AA BB BB BB BB 1712860 4 SNP_A- rs2199219 63012534 63.012534 0.321 AB AB AA AA AA AA BB BB AA AB 1706808 4 SNP_A- rs7674285 65578799 65.578799 0.536 BB BB BB BB BB BB AA AA AA AA 1657186 4 SNP_A- rs1450036 67486005 67.486005 0.619 AA AA AA AA AB AB AA AA AA AB 1701798 4 SNP_A- rs2736466 70507268 70.507268 0.679 AB AB AA AA AA AA AA AA AB AB 1734479 4 SNP_A- rs3775745 71293834 71.293834 0.536 BB BB AA AA AA AA AA AA AB AB 1645045 4 SNP_A- rs7678694 75663264 75.663264 0.476 BB BB AA AA AA AA BB BB AA AA 1741102 4 SNP_A- rs925454 77604654 77.604654 0.595 AA AA AA AA AB AB AA AA AB AB 1670999 4 SNP_A- rs2703134 78171011 78.171011 0.691 AB AB AA AA AA AA AA AB AA AA 1738063 4 SNP_A- rs10518188 79483184 79.483184 0.405 AB AB BB BB BB BB BB AB BB BB 1654306 4 SNP_A- rs2119421 80807501 80.807501 0.714 AA AA AA AA AA AA AA AA BB BB 1661108 4 SNP_A- rs9307787 83047673 83.047673 0.75 AA AA AA AA AA AA AA AA AA AA 1703940 4 SNP_A- rs6813014 84235884 84.235884 0.548 AB AB AA AA BB AB BB BB AA AA 1650367 4 SNP_A- rs10516708 85194717 85.194717 0.643 AA AA AA AA AA AA AA AA AB AB 1752998 4 SNP_A- rs10516739 86522131 86.522131 0.655 AB AB BB BB AA AB AA AA AB AB 1669642 4 SNP_A- rs10516760 87083328 87.083328 0.25 BB BB BB BB BB BB AA AA AA AA 1725569 4 SNP_A- rs4693803 88425710 88.42571 0.5 AA AA AA AA BB AB AA AA AB AB 1732366 4 SNP_A- rs10516796 89213912 89.213912 0.634 AA AA AA AA AA AA BB AB BB BB 1657663 4 SNP_A- rs1903002 90098072 90.098072 0.464 BB BB AA AA BB AB AA AA AB AB 1718322 4 SNP_A- rs7693500 90643667 90.643667 0.691 AA AA AA AA AA AA AA AB BB BB 1659867 4 SNP_A- rs4694023 91613152 91.613152 0.393 AB AB BB BB BB BB AA AA AB AB 1705800 4 SNP_A- rs7696847 92155216 92.155216 0.714 AA AA BB BB AA AB AA AB AB AB 1757446 4 SNP_A- rs6827937 94157783 94.157783 0.452 BB BB BB BB BB BB AA AB AB AB 1749382 4 SNP_A- rs10516919 94713877 94.713877 0.667 AB AB AA AA AA AA AA AA AA AA 1727842 4 SNP_A- rs1048627 95944765 95.944765 0.595 AA AA AA AA BB AB AA AA AA AA 1745861 4 SNP_A- rs1384869 96613355 96.613355 0.274 BB BB AA AA AA AA BB AB BB BB 1683945 4 SNP_A- rs6853079 99800789 99.800789 0.286 BB BB BB BB BB BB BB BB AA AA 1756011 4 SNP_A- rs1230164 100343201 100.343201 0.274 AB AB BB BB AA AB BB AB BB BB 1703546 4 SNP_A- rs238486 103377982 103.377982 0.595 AA AA AA AA BB AB AA AA BB BB 1740940 4 SNP_A- rs227284 103964838 103.964838 0.655 AA AA AA AA BB AB BB BB AB AB 1684917 4 SNP_A- rs445761 104804695 104.804695 0.679 AB AB AA AA AA AA AA AA AA AA 1753948 4 SNP_A- rs2866685 105649408 105.649408 0.5 BB BB AA AA BB AB AB AB AB AB 1747884 4 SNP_A- rs1873361 106282703 106.282703 0.345 BB BB AA AA AA AA BB BB BB BB 1720092 4 SNP_A- rs715706 106873632 106.873632 0.286 BB BB BB BB BB BB BB BB AA AA 1721929 4 SNP_A- rs1468221 108745388 108.745388 0.31 AA AA BB BB BB BB AB AB BB BB 1662125 4 SNP_A- rs7654940 110143969 110.143969 0.524 AB AB BB BB AA AB AB AB AB AB 1642856 4 SNP_A- rs6841595 113711446 113.711446 0.317 BB BB BB BB BB BB BB BB AB AB 1686749 4 SNP_A- rs10516593 114436416 114.436416 0.524 BB BB BB BB AB AB AA AA AA AA 1736814 4 SNP_A- rs998359 116228635 116.228635 0.441 BB BB BB BB BB BB BB BB AB AB 1732667 4 SNP_A- rs292910 117406405 117.406405 0.607 AA AA AA AA AA AA AA AA AB AB 1671469 4 SNP_A- rs2125710 118964366 118.964366 0.25 BB BB BB BB BB BB BB BB AB AB 1696003 4 SNP_A- rs10518293 119688966 119.688966 0.691 AA AA BB BB BB BB AA AA AB AB 1695754 4 SNP_A- rs10518336 120880537 120.880537 0.536 AA AA BB BB AB AB BB BB AB AB 1745189 4 SNP_A- rs2036696 121573324 121.573324 0.667 AA AA BB BB BB BB BB BB AB AB 1648947 4 SNP_A- rs998327 122441717 122.441717 0.726 AA AA AA AA AB AB AA AA AA AA 1702984 4 SNP_A- rs4833836 123858274 123.858274 0.381 BB BB BB BB BB BB AB AB BB BB 1733111 4 SNP_A- rs444646 124370464 124.370464 0.548 AB AB AA AA AB AB BB BB AA AA 1643743 4 SNP_A- rs10518307 125084153 125.084153 0.262 AB AB BB BB BB BB BB BB BB BB 1746251 4 SNP_A- rs7682791 125875709 125.875709 0.631 AA AA AA AA AA AA AA AA BB BB 1648121 4 SNP_A- rs953211 126708654 126.708654 0.488 AB AB AA AA AA AA AB AB AA AA 1699868 4 SNP_A- rs4834083 127331109 127.331109 0.726 AA AA BB BB AB AB AA AA AB AB 1706772 4 SNP_A- rs318510 130173248 130.173248 0.571 AA AA BB BB BB BB AA AA AA AA 1653649 4 SNP_A- rs2969001 131140397 131.140397 0.643 AA AA AA AA AA AA AB AB AA AA 1655974 4 SNP_A- rs6846560 131988611 131.988611 0.357 BB BB BB BB BB BB AB AB BB AB 1694056 4 SNP_A- rs10518609 133609659 133.609659 0.658 AA AA BB BB BB BB AA AA AA AA 1719820 4 SNP_A- rs9307688 134441091 134.441091 0.726 BB BB AA AA AA AA AA AA BB AB 1721507 4 SNP_A- rs9307703 135374277 135.374277 0.321 BB BB BB BB BB BB AB AB BB BB 1716218 4 SNP_A- rs6535037 135968531 135.968531 0.691 AB AB AA AA AA AA AB AB AA AB 1719154 4 SNP_A- rs10519369 137048788 137.048788 0.619 AB AB AA AB AB AB AB AB AA AB 1718316 4 SNP_A- rs7692053 138020209 138.020209 0.631 AB AB BB BB BB BB AA AA AA AA 1643751 4 SNP_A- rs1376088 139852726 139.852726 0.357 BB BB BB BB BB BB AB AB BB BB 1712218 4 SNP_A- rs10519540 141950175 141.950175 0.429 AB AB BB BB BB BB AA AB AA AB 1688437 4 SNP_A- rs2062597 143153292 143.153292 0.61 AA AA AA AA AA AA BB BB AA AA 1669152 4 SNP_A- rs331963 144031456 144.031456 0.369 AA AA AB AB AB AB AA AA BB AB 1651007 4 SNP_A- rs789351 146225975 146.225975 0.679 BB BB AA AA AA AA AA AA AA AA 1705078 4 SNP_A- rs10519824 148107681 148.107681 0.658 AA AA AA AA AA AA AA AA AA AA 1700284 4 SNP_A- rs6810951 149441717 149.441717 0.333 BB BB AB AB AB AB BB AB BB BB 1655082 4 SNP_A- rs10489053 150276232 150.276232 0.643 AB AB AA AA AA AA AA AA AA AB 1650743 4 SNP_A- rs991529 151852936 151.852936 0.488 BB BB BB BB BB BB AA AA AA AA 1695870 4 SNP_A- rs361101 153131731 153.131731 0.631 BB BB AB AB AA AB AA AA BB AB 1750768 4 SNP_A- rs7662116 154375569 154.375569 0.691 AA AA AA AA AA AA BB AB AA AA 1694614 4 SNP_A- rs1125228 155126657 155.126657 0.738 AA AA BB BB BB BB BB BB BB BB 1651497 4 SNP_A- rs6536240 158751762 158.751762 0.381 AB AB BB BB BB BB AB AB BB AB 1732214 4 SNP_A- rs7678486 159498426 159.498426 0.25 BB BB BB BB BB BB BB BB AA AB 1721547 4 SNP_A- rs7665879 160305968 160.305968 0.31 AB AB BB BB BB BB AB AB BB AB 1695472 4 SNP_A- rs6856295 160845965 160.845965 0.595 BB BB AB AB AA AB AB AB AA AA 1653797 4 SNP_A- rs9308000 161359479 161.359479 0.655 AA AA AA AA AA AA AA AA AA AA 1705238 4 SNP_A- rs195894 163885040 163.88504 0.366 BB BB AB AB BB AB BB BB BB BB 1679349 4 SNP_A- rs4057797 164602658 164.602658 0.381 BB BB AA AA AA AA BB BB BB AB 1719176 4 SNP_A- rs4404502 165513434 165.513434 0.738 AA AA AA AA AA AA AB AB AA AB 1657975 4 SNP_A- rs4691246 167437606 167.437606 0.402 BB BB AB AB AA AB AB AB BB BB 1669824 4 SNP_A- rs7435411 169800826 169.800826 0.524 BB BB AB AB BB AB AB AB AA AB 1701924 4 SNP_A- rs13212 170689681 170.689681 0.274 BB BB BB BB BB BB BB BB BB BB 1673825 4 SNP_A- rs449424 171884933 171.884933 0.631 AA AA AB AB AA AB AA AA AA AA 1736222 4 SNP_A- rs1485870 173125585 173.125585 0.744 BB AB AA AA AA AA AA AA AA AA 1745583 4 SNP_A- rs10520252 174445689 174.445689 0.619 AA AA AB AB AA AB AB AB AA AA 1717120 4 SNP_A- rs10520282 175752491 175.752491 0.738 AA AA AB AB AA AB AA AA AA AA 1714548 4 SNP_A- rs393279 177816548 177.816548 0.488 AA AA BB BB BB BB BB BB BB AB 1662283 4 SNP_A- rs10520383 178915654 178.915654 0.357 BB BB BB BB BB BB AB AB BB AB 1675843 4 SNP_A- rs2706012 179727204 179.727204 0.718 AA AA AA AA AA AA AA AA BB AB 1737632 4 SNP_A- rs10520430 180911694 180.911694 0.679 AA AA AB AB AA AB AA AA BB BB 1720252 4 SNP_A- rs7667245 181665445 181.665445 0.695 AA AB AB AB BB AB AA AA AA AA 1644751 4 SNP_A- rs10520479 182667862 182.667862 0.702 AA AA AA AA AA AA BB BB AA AA 1649573 4 SNP_A- rs10520518 183365702 183.365702 0.369 BB AB AB AB BB AB AA AA BB BB 1689263 4 SNP_A- rs830838 187288258 187.288258 0.333 BB AB BB AB AB AB BB BB AB AB 1738928 4 SNP_A- rs1280100 187913282 187.913282 0.345 AA AB AA AA AA AA AB AB BB BB 1713082 4 SNP_A- rs1505509 188945352 188.945352 0.345 AA AB BB BB BB BB AA AA BB BB 1654608 4 SNP_A- rs2376743 189829781 189.829781 0.679 AA AA AA AA AA AA AB AB AB AB 1680395 5 SNP_A- rs10512651 1816661 1.816661 0.405 BB BB AB AB BB AB AA AA AB AB 1689409 5 SNP_A- rs1445862 3675257 3.675257 0.25 BB BB AB AB BB AB BB AB BB BB 1642488 5 SNP_A- rs272190 5103830 5.10383 0.25 AB AB AA AA AA AA AB AB BB BB 1651781 5 SNP_A- rs2560294 5619114 5.619114 0.726 AA AA AA AA AA AA AB AB AB AB 1663379 5 SNP_A- rs10512858 6486915 6.486915 0.613 BB BB AB AB AB AB AA AA AB AB 1643560 5 SNP_A- rs4629562 7847326 7.847326 0.619 AB AB AB AB AB AB AA AA AB AB 1651637 5 SNP_A- rs9313236 8348429 8.348429 0.25 BB BB BB BB BB BB BB BB BB BB 1693207 5 SNP_A- rs12515692 9883408 9.883408 0.357 AB AB BB BB BB BB BB BB AB AB 1744546 5 SNP_A- rs2937513 11007551 11.007551 0.619 AB AB BB BB BB BB AB AB BB BB 1721268 5 SNP_A- rs173671 12217918 12.217918 0.476 AB AB AB BB AB AB AB AB AA AA 1702624 5 SNP_A- rs1476154 13000353 13.000353 0.691 BB BB AA AA AA AA AA AA AA AA 1695478 5 SNP_A- rs3734108 13806984 13.806984 0.56 AA AA AB BB AB AB AB AB AB AB 1757294 5 SNP_A- rs2938832 15817866 15.817866 0.25 AB AB BB BB BB BB AB AB AB AB 1696687 5 SNP_A- rs585991 17246633 17.246633 0.274 BB BB AA AA AA AA BB BB BB BB 1713461 5 SNP_A- rs1394215 18359538 18.359538 0.548 BB BB AA AA AA AA BB BB AA AA 1661473 5 SNP_A- rs2942296 19421031 19.421031 0.429 BB BB AB AA AB AB AB AB AA AA 1646961 5 SNP_A- rs248202 21159137 21.159137 0.274 BB BB BB BB BB BB AB AB AB AB 1698916 5 SNP_A- rs7705523 23659025 23.659025 0.714 AA AA AB BB AB AB AA AA AB AB 1757332 5 SNP_A- rs1995599 24552793 24.552793 0.548 AB AB AB BB AB AB BB BB AB AB 1672683 5 SNP_A- rs9293241 26606178 26.606178 0.56 AB AB AA AA AA AA AB AB AB AB 1660984 5 SNP_A- rs921469 29989233 29.989233 0.583 AB AB AA AA AA AA AA AA AB AB 1704664 5 SNP_A- rs1921111 30906615 30.906615 0.405 BB BB BB BB BB BB AA AA AB AB 1644843 5 SNP_A- rs893551 33493407 33.493407 0.607 AA AA AA AA AA AA AA AA AA AA 1678791 5 SNP_A- rs716302 35846025 35.846025 0.357 AB AB AA AA AA AA AA AB BB BB 1724049 5 SNP_A- rs159751 37035755 37.035755 0.464 AA AA BB BB BB BB BB BB AA AA 1703432 5 SNP_A- rs4072686 38003109 38.003109 0.405 AB AB BB BB BB BB BB BB BB AB 1645375 5 SNP_A- rs675502 39878266 39.878266 0.679 AB AB BB BB AA AB AA AA AA AA 1719252 5 SNP_A- rs1697938 40890439 40.890439 0.441 AA AA AA AA AA AA BB BB AA AA 1685613 5 SNP_A- rs276278 42016012 42.016012 0.298 BB BB BB BB BB BB BB BB BB AB 1731232 5 SNP_A- rs1072746 43646445 43.646445 0.441 AA AB BB BB BB BB BB AB AA AB 1694450 5 SNP_A- rs2404958 50098792 50.098792 0.619 AA AB AA AA BB AB AA AA AA AA 1675759 5 SNP_A- rs9283709 51510492 51.510492 0.595 BB AB BB BB AB AB AA AA BB BB 1723309 5 SNP_A- rs10512988 52085030 52.08503 0.357 BB AB AA AA AA AA BB AB AA AA 1728968 5 SNP_A- rs9292039 53454075 53.454075 0.268 BB BB BB BB BB BB AA AB BB AB 1746984 5 SNP_A- rs6450270 54287290 54.28729 0.714 AA AA AA AA AA AA BB AB AA AA 1697874 5 SNP_A- rs889310 56000924 56.000924 0.476 BB BB AB AA AB AB BB AB AA AB 1684501 5 SNP_A- rs2539731 57109292 57.109292 0.475 BB BB BB BB BB BB AB AB BB BB 1673657 5 SNP_A- rs9292159 57677129 57.677129 0.31 BB BB BB BB BB BB BB BB BB BB 1716782 5 SNP_A- rs9292180 58192447 58.192447 0.25 BB BB AB AA AB AB BB BB AA AB 1724117 5 SNP_A- rs10514860 58859777 58.859777 0.726 AA AA AA AA AA AA AA AA AA AA 1755537 5 SNP_A- rs6859376 59471964 59.471964 0.56 AA AB BB BB BB BB AB AB BB BB 1653871 5 SNP_A- rs159375 60469024 60.469024 0.691 AA AA AA AA AA AA BB BB BB AB 1653455 5 SNP_A- rs356598 63380121 63.380121 0.631 BB BB AA AA AA AA BB BB BB AB 1682537 5 SNP_A- rs7704890 66151331 66.151331 0.357 AA AB BB BB BB BB BB BB AA AB 1755307 5 SNP_A- rs6858907 67817289 67.817289 0.417 BB BB AB BB AB AB BB BB BB BB 1671457 5 SNP_A- rs1600073 74472493 74.472493 0.61 BB BB AB BB AB AB AB AB AA AA 1654744 5 SNP_A- rs10514059 75460983 75.460983 0.658 AA AA AA AA AA AA AA AA BB BB 1653531 5 SNP_A- rs2972341 76504599 76.504599 0.536 AB AB AA AA AA AA AB AB AA AB 1720510 5 SNP_A- rs949645 78478278 78.478278 0.564 AA AA AB BB AB AB BB BB AA AB 1682839 5 SNP_A- rs264986 79206180 79.20618 0.31 BB BB BB BB BB BB BB BB BB BB 1747624 5 SNP_A- rs964102 80843469 80.843469 0.679 AB AB AA AA AA AA AB AB BB BB 1730614 5 SNP_A- rs10514249 82540612 82.540612 0.56 AA AA AA AA AA AA BB AB BB BB 1732246 5 SNP_A- rs4639197 83381853 83.381853 0.25 BB BB BB BB BB BB BB BB BB BB 1729977 5 SNP_A- rs323744 86861304 86.861304 0.5 BB BB AA AA AA AA BB AB AA AB 1742238 5 SNP_A- rs819344 89093506 89.093506 0.463 AA AA AB BB AA AB BB AB BB BB 1751644 5 SNP_A- rs2935499 89626568 89.626568 0.548 AA AA BB BB BB BB AA AB AB AB 1690642 5 SNP_A- rs52308 90817903 90.817903 0.512 BB BB AA AA AA AA BB BB AA AA 1744488 5 SNP_A- rs248339 95229134 95.229134 0.643 AA AA BB BB BB BB BB AB AB AB 1670907 5 SNP_A- rs31248 96040439 96.040439 0.275 BB BB AB BB AA AB AA AB BB BB 1729028 5 SNP_A- rs10515273 97821155 97.821155 0.75 AA AA AA AA AA AA AA AA AA AA 1657092 5 SNP_A- rs2887526 98552712 98.552712 0.667 AB AB AA AA AA AA AA AA AA AA 1643346 5 SNP_A- rs2369754 99184261 99.184261 0.488 AA AA AB BB AA AB AA AB BB BB 1722905 5 SNP_A- rs1477625 101358141 101.358141 0.271 BB BB BB BB BB BB BB BB BB BB 1664073 5 SNP_A- rs9327861 101895776 101.895776 0.655 BB BB AA AA AA AA BB AB AA AA 1742802 5 SNP_A- rs39984 102625191 102.625191 0.31 AA AA BB BB BB BB AA AA BB BB 1745283 5 SNP_A- rs10515355 103975436 103.975436 0.738 AB AB AA AA AA AA BB BB AA AA 1734843 5 SNP_A- rs4957531 106511277 106.511277 0.463 AA AA AA AA AA AA AA AB AB AB 1730932 5 SNP_A- rs245243 109258634 109.258634 0.714 AB AB AB AA BB AB AA AB AA AA 1757418 5 SNP_A- rs10491424 110481705 110.481705 0.56 BB BB AB AB BB AB AA AA AA AA 1646761 5 SNP_A- rs1213404 111130917 111.130917 0.35 BB BB BB BB BB BB BB AB BB BB 1691719 5 SNP_A- rs971517 112050154 112.050154 0.476 AA AA AA AA AA AA AA AA AA AA 1747768 5 SNP_A- rs10519378 113555966 113.555966 0.738 AA AA AA AA AA AA AA AA AA AA 1726679 5 SNP_A- rs2546480 114841054 114.841054 0.452 AB AB BB BB BB BB BB BB AB AB 1738592 5 SNP_A- rs2662458 115402242 115.402242 0.655 AB AB AB AB BB AB AA AA AB AB 1708792 5 SNP_A- rs1027292 116078486 116.078486 0.548 BB BB AA AA AA AA AA AA AB AB 1720512 5 SNP_A- rs1504978 118638459 118.638459 0.655 AB AB AB AB AA AB AA AA AA AA 1701708 5 SNP_A- rs10519615 119189176 119.189176 0.643 BB BB BB BB BB BB AB AB AB AB 1689317 5 SNP_A- rs6897147 119692229 119.692229 0.691 AA AA AA AA AA AA BB BB AA AA 1751260 5 SNP_A- rs161011 123703275 123.703275 0.286 AA AA AB AB AB AB BB BB BB BB 1654688 5 SNP_A- rs7716491 124265772 124.265772 0.738 AB AB AA AA AA AA AB AB AB AB 1699578 5 SNP_A- rs1826263 124839517 124.839517 0.571 BB BB AA AA AA AA AB AB AB AB 1703238 5 SNP_A- rs964185 125631547 125.631547 0.345 AA AA AB AB AB AB BB BB AB AB 1715428 5 SNP_A- rs1345663 126678081 126.678081 0.31 BB AB AA AA AA AA AA AA AA AA 1751090 5 SNP_A- rs9327460 127338947 127.338947 0.524 AA AA AA AA AA AA AB AB BB BB 1694738 5 SNP_A- rs1181962 128414700 128.4147 0.333 BB BB AA AA AA AA AB AB BB BB 1658519 5 SNP_A- rs25810 129015788 129.015788 0.595 AA AA AA AA AA AA AA AA AB AB 1677377 5 SNP_A- rs10520083 129967905 129.967905 0.345 AA AA AB AB AB AB AB AB AB AB 1673843 5 SNP_A- rs9327673 133230970 133.23097 0.286 BB BB BB BB BB BB AB AB AB AB 1705560 5 SNP_A- rs10515473 134961986 134.961986 0.714 BB AB AA AA AA AA AA AA AB AB 1662391 5 SNP_A- rs10515481 136007946 136.007946 0.536 BB AB AA AA AA AA AA AA BB BB 1707797 5 SNP_A- rs1560930 136590879 136.590879 0.537 BB BB AA AA AA AA AA AA AB AB 1720076 5 SNP_A- rs288019 138219292 138.219292 0.39 BB BB AB AB AB AB BB AB BB BB 1697724 5 SNP_A- rs2336977 139130436 139.130436 0.61 AA AB AB AB AB AB BB AB AA AA 1707038 5 SNP_A- rs6860077 139725338 139.725338 0.31 BB AB BB BB BB BB BB BB AB AB 1703312 5 SNP_A- rs246002 140321288 140.321288 0.5 BB BB AB AB AB AB AA AB AA AA 1742086 5 SNP_A- rs32927 141102251 141.102251 0.298 AA AB BB BB BB BB BB AB AB AB 1730974 5 SNP_A- rs997833 141815738 141.815738 0.286 AA AB AB AB AB AB BB BB BB BB 1736416 5 SNP_A- rs325227 143131067 143.131067 0.31 AA AB AA AA AA AA AA AA BB BB 1722681 5 SNP_A- rs10515600 147316068 147.316068 0.548 AB AB AB AB AB AB AA AA BB BB 1749482 5 SNP_A- rs185021 148147283 148.147283 0.524 AB AB AB AB AB AB BB BB AA AA 1716760 5 SNP_A- rs10515632 149082624 149.082624 0.333 BB BB AB AB AB AB BB AB AA AA 1642124 5 SNP_A- rs1277464 150234035 150.234035 0.354 BB BB AB AB AB AB AB AB BB BB 1737743 5 SNP_A- rs2304054 150923278 150.923278 0.548 AB AB AA AA AA AA BB BB AB AB 1678329 5 SNP_A- rs10515686 152529312 152.529312 0.619 AA AA AB AB AB AB BB AB AB AB 1649583 5 SNP_A- rs4129128 153102070 153.10207 0.321 AB AB AA AA AA AA BB AB BB BB 1652471 5 SNP_A- rs991314 154438135 154.438135 0.744 AA AA AA AA AA AA AA AA AA AA 1700286 5 SNP_A- rs2569031 155177249 155.177249 0.488 AB AB AB AB AB AB AA AA AB AB 1752802 5 SNP_A- rs873343 157106698 157.106698 0.25 AB AB AB AB AB AB AA AA BB BB 1706578 5 SNP_A- rs9313777 157878177 157.878177 0.75 AA AA AA AA AA AA AA AA AB AB 1757398 5 SNP_A- rs10515781 158633942 158.633942 0.321 AB AB AB AB AB AB BB BB BB BB 1736540 5 SNP_A- rs411005 160517741 160.517741 0.476 AB AB AA AA AA AA BB BB AA AA 1647073 5 SNP_A- rs2170901 161840216 161.840216 0.429 BB BB AB AB AB AB BB BB AA AA 1724235 5 SNP_A- rs300238 162682948 162.682948 0.452 AB AB AB AB AB AB AA AA AB AB 1754048 5 SNP_A- rs158295 163217790 163.21779 0.25 BB BB BB BB BB BB AA AB BB BB 1745987 5 SNP_A- rs6869856 166017651 166.017651 0.412 AA AA AA AA AA AA AA AA AA AA 1720394 5 SNP_A- rs1911557 169681232 169.681232 0.25 BB BB BB BB BB BB AB AB AB AB 1687531 5 SNP_A- rs10516089 171083836 171.083836 0.726 AB AB AA AA AA AA AB AB AA AA 1749300 5 SNP_A- rs1909706 173644330 173.64433 0.707 AA AA AA AA AA AA AA AA AA AA 1665975 5 SNP_A- rs965017 174509108 174.509108 0.548 AB AB AB AB BB AB AA AA AA AA 1724885 5 SNP_A- rs1071882 178068646 178.068646 0.702 AA AA AB AB AA AB AA AA AA AA 1644515 5 SNP_A- rs2892344 180297919 180.297919 0.536 AB AB BB BB BB BB BB BB AB AB 1748220 6 SNP_A- rs3765437 508013 0.508013 0.536 AA AA BB BB BB BB BB BB AA AA 1732501 6 SNP_A- rs238073 1192930 1.19293 0.381 AA AA BB BB BB BB AA AA AB AB 1728682 6 SNP_A- rs6919059 1729095 1.729095 0.691 AA AB AA AA AA AA AB AB AA AA 1747718 6 SNP_A- rs2326366 3923256 3.923256 0.417 BB AB AA AB AA AB BB BB AB AB 1723553 6 SNP_A- rs10484314 5593086 5.593086 0.333 BB BB BB BB BB BB BB BB AB AB 1747058 6 SNP_A- rs3851514 6219569 6.219569 0.75 BB BB BB AB BB AB AB AB AA AA 1673883 6 SNP_A- rs267202 7799235 7.799235 0.619 AB AB AA AB AA AB AA AA AA AA 1737825 6 SNP_A- rs1543731 8355978 8.355978 0.346 AA AA AA AB AA AB BB BB BB BB 1680945 6 SNP_A- rs9296701 9687981 9.687981 0.536 AB AB BB BB BB BB BB BB AB AB 1702006 6 SNP_A- rs4512212 10379387 10.379387 0.464 BB BB AA AB AA AB BB BB BB BB 1715186 6 SNP_A- rs2182335 11324963 11.324963 0.714 AA AA AA AB AA AB BB BB AA AA 1680453 6 SNP_A- rs2841555 13574809 13.574809 0.655 AA AA AA AA AA AA AB AB AB AB 1690060 6 SNP_A- rs2237166 16755137 16.755137 0.536 AB AB AB AB BB AB BB BB AA AA 1646375 6 SNP_A- rs2147211 17898170 17.89817 0.714 AA AA AA AA AA AA AB AB AB AB 1744270 6 SNP_A- rs9297090 18873893 18.873893 0.571 AB AB AA AA AA AA AA AA AB AB 1679405 6 SNP_A- rs971623 20437442 20.437442 0.405 AB AB BB BB BB BB AB AB BB BB 1717924 6 SNP_A- rs10485012 22715005 22.715005 0.595 AB AB AB AB AB AB BB BB AA AA 1749068 6 SNP_A- rs2022330 23554534 23.554534 0.667 AB AB AB AB AB AB AB AB AB AB 1754953 6 SNP_A- rs499466 24069410 24.06941 0.5 AB AB BB BB BB BB AB AB AA AA 1698352 6 SNP_A- rs9295755 28141153 28.141153 0.25 BB BB BB BB BB BB AB AB BB BB 1682833 6 SNP_A- rs2747430 29756485 29.756485 0.702 AB AB AA AA AA AA AA AA AA AA 1656688 6 SNP_A- rs2395173 32512837 32.512837 0.691 AB AB AA AA AA AA AB AB AA AA 1715492 6 SNP_A- rs9296266 38990614 38.990614 0.573 AA AA AB AB AB AB AA AA AA AB 1722893 6 SNP_A- rs2395743 40400147 40.400147 0.488 BB AB BB BB BB BB BB BB BB AB 1724965 6 SNP_A- rs3804281 41853967 41.853967 0.429 BB BB AB AB AB AB AA AB BB BB 1757782 6 SNP_A- rs3763234 42725939 42.725939 0.298 AA AB BB BB BB BB BB BB BB AB 1700088 6 SNP_A- rs525043 44511878 44.511878 0.25 BB BB BB BB BB BB BB BB BB AB 1748380 6 SNP_A- rs9296453 45335410 45.33541 0.429 AA AB BB BB BB BB AA AA AA AA 1685295 6 SNP_A- rs9296468 45876662 45.876662 0.726 AA AA AB AB AB AB AA AA AA AA 1708722 6 SNP_A- rs10498767 46471516 46.471516 0.441 BB AB BB BB BB BB BB BB BB AB 1736458 6 SNP_A- rs9296547 47474339 47.474339 0.643 AA AA BB BB BB BB BB AB AA AA 1642956 6 SNP_A- rs2089505 48229201 48.229201 0.643 AA AA BB BB BB BB BB BB AA AA 1742558 6 SNP_A- rs504213 49411897 49.411897 0.607 AA AA AA AA AA AA AA AA BB AB 1738582 6 SNP_A- rs10484664 51124482 51.124482 0.256 BB BB BB BB BB BB BB BB BB AB 1658085 6 SNP_A- rs913098 51750772 51.750772 0.667 AA AB AA AA AA AA AA AA AA AA 1723157 6 SNP_A- rs509946 52411949 52.411949 0.369 BB BB AA BB AA AB BB AB BB BB 1726221 6 SNP_A- rs10484785 53457958 53.457958 0.476 BB AB BB BB BB BB AA AB BB BB 1717116 6 SNP_A- rs1393779 54808762 54.808762 0.464 AA AB AA AA AA AA BB BB BB BB 1717814 6 SNP_A- rs1925179 56129171 56.129171 0.655 AA AA AA AA AA AA AA AA AA AB 1693069 6 SNP_A- rs6934928 58422082 58.422082 0.714 BB AB AA AA AA AA BB AB AA AA 1664153 6 SNP_A- rs565795 62597708 62.597708 0.61 AA AA AA AA AA AA BB BB AA AB 1682123 6 SNP_A- rs9293849 63255396 63.255396 0.333 BB BB BB AA BB AB AA AB BB AB 1692597 6 SNP_A- rs9294630 65677619 65.677619 0.702 AA AB AA AA AA AA AA AA AA AB 1729072 6 SNP_A- rs2502270 67886666 67.886666 0.488 AA AA AA AA AA AA AB AB AA AA 1685655 6 SNP_A- rs4707479 68787830 68.78783 0.286 AA AA BB BB BB BB BB BB BB AB 1744006 6 SNP_A- rs579588 69639537 69.639537 0.714 AA AA BB AA BB AB AB AB BB AB 1683273 6 SNP_A- rs591809 72270133 72.270133 0.524 BB BB BB BB BB BB BB BB AA AB 1659091 6 SNP_A- rs959369 74620278 74.620278 0.607 AA AA AA AA AA AA AA AA AA AB 1660794 6 SNP_A- rs1575856 76774483 76.774483 0.262 BB BB BB BB BB BB BB BB BB AB 1656648 6 SNP_A- rs1457947 77533004 77.533004 0.619 AA AA AA AA AA AA AB AB AA AA 1675424 6 SNP_A- rs236225 79172654 79.172654 0.744 AA AA AA AA AA AA AB AB AA AA 1657250 6 SNP_A- rs239500 80761863 80.761863 0.595 AB AB AA AA AA AA AB AB BB BB 1646741 6 SNP_A- rs310387 81832380 81.83238 0.56 AA AA AA AA AA AA AA AA BB AB 1733643 6 SNP_A- rs2323435 82365338 82.365338 0.417 BB BB BB BB BB BB BB BB BB BB 1684117 6 SNP_A- rs958568 83211843 83.211843 0.417 BB BB BB BB BB BB AB AB BB AB 1749278 6 SNP_A- rs6938512 85412591 85.412591 0.476 AB AB BB BB BB BB AA AA BB BB 1737476 6 SNP_A- rs3966882 85938190 85.93819 0.46 BB BB BB AA AB AB AB AB BB BB 1658179 6 SNP_A- rs3857488 88057783 88.057783 0.548 AB AB BB BB BB BB AA AA AA AB 1704672 6 SNP_A- rs942115 90274825 90.274825 0.691 AA AA BB BB BB BB AA AA AA AA 1750678 6 SNP_A- rs1753826 91283465 91.283465 0.274 BB BB BB BB BB BB BB BB BB BB 1641794 6 SNP_A- rs427118 92400922 92.400922 0.622 AB AB AA AA AA AA AA AA AB AB 1747902 6 SNP_A- rs609590 93182864 93.182864 0.595 AB AB AA AA AA AA AB AB AB AB 1699604 6 SNP_A- rs1906966 94362973 94.362973 0.571 AA AA AA AA AA AA AB AB AB AB 1671865 6 SNP_A- rs2380218 95947837 95.947837 0.643 AB AB BB AB AB AB AB AB AA AA 1672477 6 SNP_A- rs6925466 96483564 96.483564 0.476 AB AB AA AB AB AB BB BB AB AB 1727169 6 SNP_A- rs2206094 97681917 97.681917 0.595 AB AB AA AA AA AA AB AB AB AB 1669372 6 SNP_A- rs10484477 103889951 103.889951 0.738 AA AA AA AA AA AA BB BB AB AB 1754567 6 SNP_A- rs1341123 104971433 104.971433 0.274 BB BB BB BB BB BB BB BB AB AB 1687147 6 SNP_A- rs1325421 105998201 105.998201 0.643 AB AB AA AB AB AB AA AA AA AA 1696467 6 SNP_A- rs1462145 107068713 107.068713 0.357 AB AB BB BB BB BB BB BB BB BB 1733167 6 SNP_A- rs7740028 110825873 110.825873 0.393 AA AA BB BB BB BB AA AA BB BB 1650105 6 SNP_A- rs2010315 112529340 112.52934 0.679 AA AA AA AA AA AA AA AA AA AA 1680493 6 SNP_A- rs1378682 113188320 113.18832 0.405 AA AA BB BB BB BB AA AA AB AB 1735595 6 SNP_A- rs2810160 114329403 114.329403 0.548 AA AB BB BB BB BB AB AB AB AB 1712634 6 SNP_A- rs1748168 114955404 114.955404 0.56 BB AB AA AA AA AA AB AB BB BB 1687581 6 SNP_A- rs2250263 116940700 116.9407 0.679 AA AB AA AA AA AA AA AA BB BB 1738139 6 SNP_A- rs929122 117712442 117.712442 0.726 AA AA AA AA AA AA AA AA AB AB 1729937 6 SNP_A- rs9285429 118811259 118.811259 0.619 AA AA AB AB AB AB AA AA BB BB 1661803 6 SNP_A- rs1873553 120164641 120.164641 0.429 BB BB BB BB BB BB AB AB AA AA 1665123 6 SNP_A- rs6906196 122713901 122.713901 0.369 BB BB BB BB BB BB BB BB BB BB 1679087 6 SNP_A- rs6924068 124232587 124.232587 0.345 BB BB AB AB AB AB BB BB BB BB 1707510 6 SNP_A- rs484510 125528599 125.528599 0.691 AA AA AA AA AA AA AA AA AB AB 1745119 6 SNP_A- rs9321057 126198636 126.198636 0.405 AA AB BB BB BB BB AB AB AB AB 1685381 6 SNP_A- rs270044 128228176 128.228176 0.405 AA AB BB BB BB BB BB BB AB AB 1751986 6 SNP_A- rs1508439 129073191 129.073191 0.655 AA AB AA AA AA AA AB AB AA AA 1721422 6 SNP_A- rs10484282 130107771 130.107771 0.321 BB AB BB BB BB BB AB AB BB BB 1707720 6 SNP_A- rs766967 130912718 130.912718 0.488 AA AA AB AB AB AB AB AB BB BB 1652817 6 SNP_A- rs170881 132358254 132.358254 0.655 AA AA BB BB BB BB AB AB BB BB 1653251 6 SNP_A- rs2745426 133045037 133.045037 0.287 BB BB BB BB BB BB BB BB BB BB 1723663 6 SNP_A- rs509904 133558775 133.558775 0.713 AA AB AA AA AA AA AA AA BB BB 1751418 6 SNP_A- rs6570091 137092589 137.092589 0.464 BB BB AA AA AA AA BB BB AB AB 1735812 6 SNP_A- rs662100 137931606 137.931606 0.512 AA AA AA AA AA AA BB AB AA AA 1669540 6 SNP_A- rs2473522 139472945 139.472945 0.274 BB BB AB AB AB AB AA AA BB BB 1692831 6 SNP_A- rs9321743 140005650 140.00565 0.655 AA AA AA AA AA AA AA AA AB AB 1729139 6 SNP_A- rs225710 142582952 142.582952 0.548 AB AB BB BB BB BB BB AB AA AA 1668519 6 SNP_A- rs10484804 143972461 143.972461 0.262 AB AB BB BB BB BB BB BB BB BB 1741850 6 SNP_A- rs4243477 145767511 145.767511 0.655 AA AA AA AA AA AA AA AB AB AB 1699598 6 SNP_A- rs6923545 146286436 146.286436 0.417 AB AB BB BB BB BB BB AB AB AB 1714061 6 SNP_A- rs2025157 146851348 146.851348 0.607 AA AA AA AA AA AA AA AB BB BB 1643757 6 SNP_A- rs10484677 148334072 148.334072 0.702 AA AA AA AA AA AA AA AA AB AB 1746680 6 SNP_A- rs997682 149708313 149.708313 0.293 BB BB AB AB AB AB BB AB BB BB 1674099 6 SNP_A- rs1933079 151696424 151.696424 0.464 AA AA BB BB BB BB AA AB BB BB 1742378 6 SNP_A- rs872371 153469676 153.469676 0.732 AA AA BB BB BB BB AA AB AA AA 1664463 6 SNP_A- rs612450 154306471 154.306471 0.56 AB AB AA AB AB AB AA AA AA AA 1706738 6 SNP_A- rs1980602 155248334 155.248334 0.429 AB AB AA AA AA AA BB AB AB AB 1661002 6 SNP_A- rs1391655 156092837 156.092837 0.298 BB AB BB BB BB BB BB AB BB BB 1700465 6 SNP_A- rs7770496 156806897 156.806897 0.372 AA AB AA AB AB AB AA AB AB AB 1660620 6 SNP_A- rs4709298 157885415 157.885415 0.262 BB AB BB AB AB AB AA AB AA AA 1712976 6 SNP_A- rs7753885 158892133 158.892133 0.488 BB AB AA AB AB AB BB BB BB BB 1676117 6 SNP_A- rs923459 159532261 159.532261 0.513 BB AB AA AA AA AA BB BB AB AB 1669774 6 SNP_A- rs927450 160152507 160.152507 0.583 AA AA AA AA AA AA BB AB AB AB 1659978 6 SNP_A- rs598969 160664317 160.664317 0.425 BB AB AA AA AA AA BB BB BB BB 1670969 6 SNP_A- rs6910079 164339862 164.339862 0.441 AA AB BB AB AB AB BB BB AB AB 1697554 6 SNP_A- rs907223 165179009 165.179009 0.702 AA AA AA AA AA AA AA AA AB AB 1641972 6 SNP_A- rs162293 167420582 167.420582 0.75 AA AA AA AA AA AA AA AA AA AA 1698488 7 SNP_A- rs1637750 2001052 2.001052 0.655 AA AA AA AA AA AA AB AB AA AA 1647507 7 SNP_A- rs10257982 3107838 3.107838 0.452 AA AB AA AA AA AA BB BB AB AB 1710599 7 SNP_A- rs10488360 4184450 4.18445 0.317 BB BB BB BB BB BB BB BB BB BB 1675597 7 SNP_A- rs719423 7128355 7.128355 0.667 AA AA BB BB BB BB BB BB AB AB 1712104 7 SNP_A- rs38012 7815795 7.815795 0.464 AB AB AB AB BB AB AB AB AB AB 1649251 7 SNP_A- rs10253058 10364900 10.3649 0.738 AA AA AA AA AA AA AA AA AB AB 1714013 7 SNP_A- rs7785008 10921568 10.921568 0.452 AB AB BB BB BB BB AA AA AB AB 1656052 7 SNP_A- rs10270630 11629477 11.629477 0.738 BB BB AB AB BB AB AA AA AB AB 1678735 7 SNP_A- rs1036667 12197823 12.197823 0.488 BB BB AB AB AA AB AA AA AA AA 1672885 7 SNP_A- rs2214867 13507965 13.507965 0.417 AA AA AA AA AA AA BB BB AB AB 1707354 7 SNP_A- rs7793372 14320689 14.320689 0.619 BB BB BB BB BB BB AA AA AB AB 1724539 7 SNP_A- rs1527203 15931001 15.931001 0.643 AA AA AB AB AB AB AA AA AA AA 1756798 7 SNP_A- rs706057 16577230 16.57723 0.56 AB AB AB AB AB AB AB AB AB AB 1755023 7 SNP_A- rs4721619 17291814 17.291814 0.667 AB AB AA AA AA AA AB AB AA AA 1647845 7 SNP_A- rs2731551 18050825 18.050825 0.25 BB BB AB AB AB AB BB BB BB BB 1693824 7 SNP_A- rs10486334 18974842 18.974842 0.726 BB BB AB AB AB AB BB BB AA AA 1724213 7 SNP_A- rs2248634 21065118 21.065118 0.262 BB BB BB BB BB BB AB AB BB BB 1716746 7 SNP_A- rs7781044 21636203 21.636203 0.61 AA AA AA AA AA AA AB AB AB AB 1706218 7 SNP_A- rs2286497 22701238 22.701238 0.305 BB BB AB AB AB AB AB AB AB AB 1718088 7 SNP_A- rs2521642 24200036 24.200036 0.31 BB BB BB BB BB BB AB AB BB BB 1755947 7 SNP_A- rs4275130 26366016 26.366016 0.75 AB AB AA AA AA AA AA AA AA AA 1751950 7 SNP_A- rs6953785 27237607 27.237607 0.536 AB AB AB BB AB AB AB AB AA AA 1725907 7 SNP_A- rs4498447 28177012 28.177012 0.524 BB BB AB BB AB AB BB BB AB AB 1729503 7 SNP_A- rs1859681 28699408 28.699408 0.275 BB BB BB BB BB BB BB BB BB BB 1663287 7 SNP_A- rs1476991 29293282 29.293282 0.571 AA AA AA AA AA AA AA AA AB AB 1728544 7 SNP_A- rs997349 29955684 29.955684 0.357 BB BB BB BB BB BB BB BB BB BB 1718026 7 SNP_A- rs10487729 31345524 31.345524 0.726 AA AA AA AA AA AA AB AB BB BB 1695134 7 SNP_A- rs215675 32156237 32.156237 0.286 AA AB BB BB BB BB BB BB AB AB 1694740 7 SNP_A- rs10254116 33010729 33.010729 0.631 BB AB AA AA AA AA AA AA AA AA 1717858 7 SNP_A- rs10486619 33600838 33.600838 0.262 BB BB BB BB BB BB BB BB AB AB 1678175 7 SNP_A- rs741202 35093154 35.093154 0.262 BB BB BB BB AB AB BB BB AA AA 1749566 7 SNP_A- rs4720228 36725169 36.725169 0.476 BB AB BB BB BB BB BB BB BB BB 1731924 7 SNP_A- rs2893552 37928803 37.928803 0.679 AA AA AA AA AA AA BB BB AA AA 1671783 7 SNP_A- rs4723791 38613746 38.613746 0.56 BB AB AA AA AA AA AA AA AA AA 1730217 7 SNP_A- rs10486802 39497008 39.497008 0.571 AA AA BB BB BB BB AA AA AA AA 1669882 7 SNP_A- rs7807596 40599281 40.599281 0.417 AA AA BB BB AB AB AB AB AA AA 1721388 7 SNP_A- rs384469 41099793 41.099793 0.702 AA AB AA AA AA AA AB AB AA AA 1750290 7 SNP_A- rs721273 42867596 42.867596 0.702 AA AA BB BB AB AB AA AA AA AA 1695614 7 SNP_A- rs2330918 43444472 43.444472 0.75 AA AB AA AA AA AA AA AA AA AA 1736506 7 SNP_A- rs10253161 46769632 46.769632 0.714 AA AA AA AA AA AA AA AA BB AB 1732094 7 SNP_A- rs7357251 47841498 47.841498 0.417 AA AA AA AA AA AB BB BB BB BB 1710294 7 SNP_A- rs3923511 48463293 48.463293 0.688 AA AA AA AA AA AA AB AB BB AB 1669906 7 SNP_A- rs716719 50102978 50.102978 0.262 AA AA BB BB BB BB BB BB BB BB 1748806 7 SNP_A- rs2159809 52287324 52.287324 0.39 AA AB BB BB BB BB BB BB BB BB 1646085 7 SNP_A- rs6955211 63316490 63.31649 0.464 BB AB AA AA AA AA AA AB BB BB 1655668 7 SNP_A- rs9638255 67214110 67.21411 0.655 AA AA AA AA AA AA AA AB AA AA 1695272 7 SNP_A- rs1699443 68224124 68.224124 0.583 AA AB AA AA AA AA AA AA BB AB 1673105 7 SNP_A- rs10499812 69098641 69.098641 0.333 AA AB BB BB BB BB BB BB BB BB 1667673 7 SNP_A- rs6976144 77019455 77.019455 0.5 AA AB BB BB BB BB AB AB AA AB 1757146 7 SNP_A- rs10485887 77712706 77.712706 0.548 AA AA AA AB AA AB BB BB AA AB 1741890 7 SNP_A- rs984312 78285441 78.285441 0.631 AA AA BB AB AB AB AA AA BB AB 1663217 7 SNP_A- rs3211816 79922641 79.922641 0.39 BB BB BB BB BB BB AB AB BB BB 1676663 7 SNP_A- rs3801720 81447606 81.447606 0.595 AA AB BB AB AB AB AB AB BB AB 1724625 7 SNP_A- rs1693380 82818863 82.818863 0.738 AA AA AA AA AA AA AB AB AA AA 1701440 7 SNP_A- rs10499889 84765715 84.765715 0.369 BB BB BB BB BB BB AB AB AA AA 1690947 7 SNP_A- rs1063964 87480120 87.48012 0.607 AA AA AA AA AA AA AA AA BB AB 1722683 7 SNP_A- rs7799830 88761617 88.761617 0.429 BB BB AA AA AA AA AB AB AA AA 1697794 7 SNP_A- rs3802029 90126750 90.12675 0.595 AB AB AA AB AB AB BB BB BB AB 1692549 7 SNP_A- rs1468180 92759526 92.759526 0.441 BB BB AA AA AA AA AA AA AA AB 1721485 7 SNP_A- rs6465448 94217939 94.217939 0.548 AB AB BB BB BB BB AA AA BB AB 1705566 7 SNP_A- rs1403179 96113755 96.113755 0.75 AB AB AA AA AA AA AB AB AA AA 1644895 7 SNP_A- rs7779090 96790254 96.790254 0.345 BB BB BB BB BB BB AB AB BB AB 1698924 7 SNP_A- rs2572009 99133656 99.133656 0.524 AA AA BB AB AB AB AA AA AA AA 1755481 7 SNP_A- rs10487284 102064226 102.064226 0.667 AA AA AA AA AA AA AA AA AA AB 1669180 7 SNP_A- rs10487162 102860400 102.8604 0.256 AB AB BB AB AB AB BB BB BB AB 1730488 7 SNP_A- rs2519681 105578447 105.578447 0.369 AB AB AA AB AB AB AB AB AA AB 1715320 7 SNP_A- rs997381 106280867 106.280867 0.524 AB AB BB BB BB BB AA AA BB BB 1657867 7 SNP_A- rs3801948 106832398 106.832398 0.643 AA AA BB AB AB AB AA AA BB AB 1703262 7 SNP_A- rs1015422 107930809 107.930809 0.298 BB AB AA AB AB AB AB AB BB AB 1687475 7 SNP_A- rs2106442 108493824 108.493824 0.476 AA AA BB BB BB BB AB AB AA AA 1643849 7 SNP_A- rs10487320 109537858 109.537858 0.619 BB AB BB BB BB BB AA AB AA AA 1688527 7 SNP_A- rs10500003 110076704 110.076704 0.702 AA AB AA AA AA AA AA AA AA AA 1641802 7 SNP_A- rs10487331 110945463 110.945463 0.726 AA AA AA AB AB AB BB AB AA AB 1740412 7 SNP_A- rs2529588 111697006 111.697006 0.726 AA AA AA AA AA AA AA AB AA AB 1745955 7 SNP_A- rs1548395 112947523 112.947523 0.298 BB BB BB BB BB BB BB AB AA AB 1724315 7 SNP_A- rs6973150 114297804 114.297804 0.45 BB BB AB AB AB AB BB BB BB BB 1719538 7 SNP_A- rs10500054 115247129 115.247129 0.5 BB BB BB BB BB BB AA AB BB BB 1736164 7 SNP_A- rs7783832 116468422 116.468422 0.298 BB BB AB AB AB AB BB AB BB AB 1733815 7 SNP_A- rs10487392 117466579 117.466579 0.488 BB BB AB AB AB AB AA AA AA AB 1676935 7 SNP_A- rs10488301 119761267 119.761267 0.262 AA AB BB BB BB BB AA AB BB BB 1647647 7 SNP_A- rs1206486 121146345 121.146345 0.441 BB BB BB BB BB BB AA AB BB BB 1655036 7 SNP_A- rs10487974 122442567 122.442567 0.381 AA AB AB AB AB AB BB BB BB BB 1643783 7 SNP_A- rs6948425 123386447 123.386447 0.476 AA AA AB AB AB AB AA AB BB AB 1690238 7 SNP_A- rs723444 124253175 124.253175 0.417 BB BB AB AB AB AB BB BB BB BB 1745008 7 SNP_A- rs2107098 124969695 124.969695 0.488 BB AB AA AA AA AA BB AB BB AB 1675675 7 SNP_A- rs2299447 125743520 125.74352 0.738 AA AA AA AA AA AA AA AA AA AA 1678693 7 SNP_A- rs6467115 126530478 126.530478 0.643 AA AA AA AA AA AA AA AA BB BB 1670675 7 SNP_A- rs10487505 127454114 127.454114 0.464 BB BB AB AB AB AB BB BB BB AB 1708033 7 SNP_A- rs10488628 127961843 127.961843 0.405 AA AB AB AB AB AB AA AA AA AB 1656498 7 SNP_A- rs7803075 130199321 130.199321 0.726 AA AA BB BB BB BB BB AB AA AA 1742598 7 SNP_A- rs1790998 133595635 133.595635 0.476 AA AA AB AB AB AB BB BB BB BB 1695048 7 SNP_A- rs2551778 134556904 134.556904 0.548 AA AA AA AA AA AA AA AA AA AA 1721434 7 SNP_A- rs10253975 135674764 135.674764 0.662 BB AB AA AA AA AA AA AA AA AA 1669022 7 SNP_A- rs2253729 139396073 139.396073 0.583 AA AB BB BB BB BB AA AA BB BB 1715624 7 SNP_A- rs1527304 141162389 141.162389 0.571 AA AA BB BB BB BB AA AA AA AB 1652925 7 SNP_A- rs6949653 143162918 143.162918 0.691 AA AB AB AB AB AB AA AA AA AA 1715016 7 SNP_A- rs4725680 144870787 144.870787 0.369 AA AB BB BB BB BB AA AA AA AA 1666637 7 SNP_A- rs10487936 145527849 145.527849 0.732 AA AA AA AA AA AA AA AA BB AB 1681703 7 SNP_A- rs10278315 146403556 146.403556 0.726 AA AA AA AA AA AA AA AA AA AA 1680321 7 SNP_A- rs1177946 147495250 147.49525 0.643 AA AA AA AA AA AA AA AA BB BB 1720798 7 SNP_A- rs1403222 149691561 149.691561 0.536 AB AB AA AA AA AA BB BB AA AB 1713513 7 SNP_A- rs306293 154243700 154.2437 0.643 AA AA BB BB BB BB AA AA AB AB 1736364 7 SNP_A- rs2301916 156473603 156.473603 0.369 AB AB AB AB BB AB BB BB AB AB 1691199 8 SNP_A- rs747351 228574 0.228574 0.369 AB AB AB AB AA AB AB AB AA AB 1659972 8 SNP_A- rs4876153 2291741 2.291741 0.691 BB BB AA AA AA AA AA AA AA AB 1725579 8 SNP_A- rs9314492 3332437 3.332437 0.476 AA AA BB BB BB BB BB BB BB AB 1651465 8 SNP_A- rs10503246 4117771 4.117771 0.714 AB AB AA AA AA AA BB AB BB AB 1695486 8 SNP_A- rs4146469 5253259 5.253259 0.441 AB AB AB AB AB AB BB AB BB BB 1650643 8 SNP_A- rs6559072 5839489 5.839489 0.679 AB AB AA AA AA AA BB BB AA AA 1714359 8 SNP_A- rs3020252 6450411 6.450411 0.634 AB AB BB BB BB BB AA AA BB AB 1660050 8 SNP_A- rs2409113 8849712 8.849712 0.72 AB AB AA AA AA AA AA AA AA AA 1757262 8 SNP_A- rs1588198 9929939 9.929939 0.655 AB AB AB AB AB AB AA AB AA AA 1680667 8 SNP_A- rs2278335 10740863 10.740863 0.702 AB AB AA AA AA AA AA AA AA AB 1679891 8 SNP_A- rs10503478 13876453 13.876453 0.607 AA AA AB AB AB AB AA AB BB BB 1715348 8 SNP_A- rs2410193 14445035 14.445035 0.738 AA AA AA AA AA AA BB BB AA AB 1659353 8 SNP_A- rs351572 16065839 16.065839 0.595 AA AA AA AA AA AA AA AA BB AB 1756484 8 SNP_A- rs7003503 17226143 17.226143 0.25 BB BB AB AA AB AB BB BB BB AB 1750990 8 SNP_A- rs7006702 19316813 19.316813 0.333 AA AB AB BB AB AB BB BB BB AB 1688509 8 SNP_A- rs2083637 19909455 19.909455 0.738 AA AB AB BB AB AB AB AB AA AA 1747706 8 SNP_A- rs2306518 22526253 22.526253 0.357 BB AB BB BB BB BB BB BB AA AA 1646595 8 SNP_A- rs10503733 23589963 23.589963 0.714 AA AB AA AA AA AA AB AB AA AA 1699334 8 SNP_A- rs2976457 24923988 24.923988 0.548 BB AB AA AA AA AA AB AB BB BB 1752532 8 SNP_A- rs10503776 25765786 25.765786 0.671 BB BB AA AA AA AA AB AB AA AA 1746191 8 SNP_A- rs10503872 30556573 30.556573 0.476 AA AB BB AA AA AB AA AA BB BB 1742962 8 SNP_A- rs10503907 32291552 32.291552 0.607 AA AA AA BB BB AB AB AB AA AB 1710298 8 SNP_A- rs1551652 34443033 34.443033 0.662 AA AA BB AA AA AB BB BB AA AA 1646333 8 SNP_A- rs10503970 34985910 34.98591 0.262 BB BB BB AA AA AB AB AB BB BB 1679337 8 SNP_A- rs581187 37119893 37.119893 0.286 BB BB AA AA AA AA BB BB BB AB 1701068 8 SNP_A- rs3935233 39307991 39.307991 0.31 AA AB BB BB BB BB BB BB BB BB 1747018 8 SNP_A- rs9298596 40431722 40.431722 0.333 AA AA BB AA AA AB BB BB BB BB 1730295 8 SNP_A- rs341817 50186153 50.186153 0.56 AA AA AA BB BB AB AA AB BB AB 1664173 8 SNP_A- rs318913 51075845 51.075845 0.262 BB BB BB BB BB BB BB BB BB BB 1712754 8 SNP_A- rs10504120 52554998 52.554998 0.726 AA AA AA AA AA AA AA AA AA AA 1716236 8 SNP_A- rs2249236 53110767 53.110767 0.286 AA AB AA BB AB AB AA AB BB BB 1645251 8 SNP_A- rs360956 54063839 54.063839 0.61 BB BB BB AA AB AB AA AA BB BB 1674928 8 SNP_A- rs7824078 55966296 55.966296 0.631 AA AA BB AA AB AB AA AA AA AB 1661925 8 SNP_A- rs2670052 57666163 57.666163 0.583 AA AB AA BB AB AB AA AA AA AA 1734483 8 SNP_A- rs9297980 58641477 58.641477 0.476 AA AA AB BB AB AB AA AB AB AB 1649879 8 SNP_A- rs7012230 62449232 62.449232 0.31 BB BB BB BB BB BB BB BB AB AB 1689109 8 SNP_A- rs874777 65147501 65.147501 0.583 BB BB BB BB BB BB AA AA AA AA 1729837 8 SNP_A- rs977467 67469418 67.469418 0.56 BB AB AA AA AA AA AA AB AB AB 1688563 8 SNP_A- rs900896 68690751 68.690751 0.702 AA AA AB AA AB AB AA AA AA AA 1656454 8 SNP_A- rs1404605 69369253 69.369253 0.585 BB AB AA AA AA AA BB BB AA AA 1673083 8 SNP_A- rs10504451 70626182 70.626182 0.524 AA AB BB BB BB BB AA AA AB AB 1673921 8 SNP_A- rs10504477 71500739 71.500739 0.487 BB AB AB AA AB AB BB AB AB AB 1660240 8 SNP_A- rs2732090 72080811 72.080811 0.5 AA AB AA AA AA AA AA AB AB AB 1698932 8 SNP_A- rs10504526 73129106 73.129106 0.548 AA AA AA AA AA AA BB AB AA AA 1710462 8 SNP_A- rs10504552 75038119 75.038119 0.286 BB BB AA AA AA AA AA AA AA AA 1684163 8 SNP_A- rs1375646 76679672 76.679672 0.321 BB BB BB BB BB BB BB BB AB AB 1673775 8 SNP_A- rs1993196 78213269 78.213269 0.583 BB BB AA AA AA AA AB AB AA AA 1753574 8 SNP_A- rs2461063 80781668 80.781668 0.631 AA AA AB BB AB AB AB AB AB AB 1713893 8 SNP_A- rs1199030 81917969 81.917969 0.357 BB BB AA AA AA AA BB BB BB BB 1650035 8 SNP_A- rs1525339 83916405 83.916405 0.738 AA AA AA AA AA AA BB BB AA AA 1709456 8 SNP_A- rs1465809 85243012 85.243012 0.25 BB BB BB BB BB BB BB BB AB AB 1747972 8 SNP_A- rs3808538 86308563 86.308563 0.738 AA AA BB BB BB BB AA AA AB AB 1690861 8 SNP_A- rs10504819 87183400 87.1834 0.369 BB BB AA AA AA AA AA AA BB BB 1642120 8 SNP_A- rs997597 88259667 88.259667 0.274 BB BB BB BB BB BB BB BB BB BB 1704458 8 SNP_A- rs10504855 88844371 88.844371 0.345 BB BB BB BB BB BB AB AB AB AB 1731702 8 SNP_A- rs160410 90717844 90.717844 0.658 AA AA BB BB BB BB AA AA AA AA 1669078 8 SNP_A- rs1818193 91886818 91.886818 0.631 AA AA AA AA AA AA AA AA AB AB 1713264 8 SNP_A- rs2245797 95329376 95.329376 0.31 BB BB BB BB BB BB AB AB AB AB 1679699 8 SNP_A- rs962451 101400186 101.400186 0.583 AA AA AA AA AA AA AA AA BB BB 1738642 8 SNP_A- rs4495397 103476369 103.476369 0.268 BB BB AA AA AA AA BB BB BB BB 1677965 8 SNP_A- rs543736 104082125 104.082125 0.619 AB AB BB BB BB BB AA AB AA AA 1718730 8 SNP_A- rs10505064 105831730 105.83173 0.345 AB AB BB BB BB BB BB AB BB BB 1724051 8 SNP_A- rs2930485 107881851 107.881851 0.607 AB AB AB BB AB AB AA AB BB BB 1691919 8 SNP_A- rs10505107 108392560 108.39256 0.619 AA AA AB AA AB AB AA AA AA AA 1652191 8 SNP_A- rs1353298 108959098 108.959098 0.321 BB BB BB BB BB BB BB BB AB AB 1756952 8 SNP_A- rs5772 110167808 110.167808 0.571 AB AB AB BB AB AB AA AB AB AB 1695466 8 SNP_A- rs10505135 111120579 111.120579 0.345 AA AA BB BB BB BB AA AB AB AB 1747370 8 SNP_A- rs10505156 112369457 112.369457 0.25 AB AB BB BB BB BB BB BB BB BB 1745327 8 SNP_A- rs10505180 113392265 113.392265 0.726 AB AB AA AA AA AA AA AA AA AA 1681911 8 SNP_A- rs2125552 113984509 113.984509 0.274 BB BB AB BB AB AB BB AB BB BB 1694542 8 SNP_A- rs9297496 114629527 114.629527 0.321 BB BB BB BB BB BB BB BB AB AB 1713409 8 SNP_A- rs7828185 116438576 116.438576 0.286 BB BB BB BB BB BB BB BB BB BB 1725803 8 SNP_A- rs10505328 119219639 119.219639 0.441 AB AB BB BB BB BB BB BB BB BB 1698988 8 SNP_A- rs3924784 121618858 121.618858 0.667 AA AA AA AA AA AA BB AB AB AB 1753414 8 SNP_A- rs17478 122793072 122.793072 0.595 AA AA AB AA AB AB BB AB AB AB 1735413 8 SNP_A- rs6470143 124219594 124.219594 0.345 BB BB BB BB BB BB BB AB AA AA 1655430 8 SNP_A- rs3909562 124803864 124.803864 0.405 AA AB AA AA AA AA BB BB AB AB 1754805 8 SNP_A- rs2382993 125770106 125.770106 0.345 BB AB BB BB BB BB BB AB BB BB 1696789 8 SNP_A- rs897153 126747483 126.747483 0.643 BB AB AB BB AB AB AA AB AB AB 1686811 8 SNP_A- rs2091933 127485749 127.485749 0.679 AA AB BB BB BB BB AA AA AB AB 1753008 8 SNP_A- rs10505486 128074016 128.074016 0.441 BB BB BB BB BB BB BB AB AB AB 1651085 8 SNP_A- rs4123791 129288419 129.288419 0.417 BB BB AA AA AA AA AA AA AA AA 1682761 8 SNP_A- rs9297775 129805894 129.805894 0.333 BB AB BB BB BB BB BB BB BB BB 1692841 8 SNP_A- rs10505545 130646449 130.646449 0.538 AA AA AB AB AB AB BB BB BB BB 1653731 8 SNP_A- rs7460225 131408555 131.408555 0.464 AA AB AB AB AB AB AA AB AA AA 1655374 8 SNP_A- rs7008202 132143592 132.143592 0.357 BB AB AA AA AA AA BB BB AB AB 1672735 8 SNP_A- rs4736424 133782292 133.782292 0.31 BB BB BB BB BB BB AA AB BB BB 1725115 8 SNP_A- rs10505607 134527931 134.527931 0.441 AA AB BB BB BB BB BB BB AB AB 1647079 8 SNP_A- rs4909801 135948341 135.948341 0.702 AA AB BB BB BB BB AA AB AA AA 1700220 8 SNP_A- rs4909582 137288153 137.288153 0.488 AA AA BB AB AB AB BB BB AA AA 1675316 8 SNP_A- rs9324439 138086052 138.086052 0.452 BB AB BB BB BB BB BB BB AB AB 1661056 8 SNP_A- rs1325053 139156386 139.156386 0.732 AA AA AA AB AB AB BB BB AA AA 1689603 8 SNP_A- rs2468705 140618265 140.618265 0.75 AB AB BB AB AB AB AA AA AA AA 1710354 9 SNP_A- rs10491691 336963 0.336963 0.655 AB AB AA AA AA AA AB AB BB BB 1643050 9 SNP_A- rs2370220 907667 0.907667 0.726 AA AA AB AB AB AB AA AA AA AA 1704718 9 SNP_A- rs7040916 2645520 2.64552 0.726 BB BB AA AA AA AA AA AA AA AA 1681445 9 SNP_A- rs1358908 3162093 3.162093 0.524 AA AA BB BB BB BB BB BB AA AA 1734535 9 SNP_A- rs1455177 3782613 3.782613 0.488 BB BB BB BB BB BB AB AB AB AB 1685961 9 SNP_A- rs10491650 5193054 5.193054 0.354 BB BB BB BB BB BB BB BB BB BB 1642494 9 SNP_A- rs1407473 7989681 7.989681 0.31 BB BB BB BB BB BB BB BB BB BB 1696419 9 SNP_A- rs1433548 8927116 8.927116 0.31 AB AB AB AB AB AB BB BB BB BB 1709516 9 SNP_A- rs1613507 9791755 9.791755 0.563 AB AB AA AA AA AA BB BB AA AA 1682679 9 SNP_A- rs10511545 10341048 10.341048 0.726 AA AA AA AA AA AA AA AA AA AA 1673445 9 SNP_A- rs4740473 11080005 11.080005 0.369 AB AB AA AA AA AA BB BB BB BB 1679185 9 SNP_A- rs1825739 11777410 11.77741 0.429 BB BB BB BB BB BB AB AB AA AB 1744452 9 SNP_A- rs1086377 12824688 12.824688 0.702 AA AA BB BB BB BB AB AB AA AA 1714556 9 SNP_A- rs7038474 13355816 13.355816 0.702 AB AB AA AA AA AA BB BB BB AB 1672461 9 SNP_A- rs10511587 13970584 13.970584 0.738 AB AB AA AA AA AA AB AB AA AA 1704214 9 SNP_A- rs4615688 14495861 14.495861 0.714 AA AA AA AA AA AA AA AA AA AA 1714243 9 SNP_A- rs10511603 15006475 15.006475 0.536 BB BB BB BB BB BB AB AB AA AA 1741448 9 SNP_A- rs1001265 17618726 17.618726 0.726 BB BB AA AA AA AA AA AA AA AA 1742240 9 SNP_A- rs7862683 18257947 18.257947 0.427 BB BB AA AA AB AB AB AB BB BB 1675206 9 SNP_A- rs7859334 20660966 20.660966 0.56 AA AA BB BB AB AB AB AB BB AB 1706426 9 SNP_A- rs871024 21793880 21.79388 0.441 AA AA AA AA AA AA AB AB BB AB 1669996 9 SNP_A- rs10511705 22537789 22.537789 0.512 BB BB BB BB AB AB AB AB BB AB 1662201 9 SNP_A- rs9298846 23216243 23.216243 0.655 AB AB AA AA AA AA AA AA AA AB 1673761 9 SNP_A- rs10511761 25602704 25.602704 0.441 BB BB BB BB BB BB AA AA AA AA 1752066 9 SNP_A- rs4978049 26131011 26.131011 0.381 AB AB BB BB BB BB BB BB BB BB 1690106 9 SNP_A- rs983863 26681668 26.681668 0.345 BB BB BB BB AB AB AA AA AA AA 1700687 9 SNP_A- rs1452357 28090846 28.090846 0.707 BB BB BB BB AB AB AA AA AA AA 1690672 9 SNP_A- rs824257 28765262 28.765262 0.655 AA AA AA AA AA AA AA AA BB BB 1693514 9 SNP_A- rs10511842 30009704 30.009704 0.25 AA AB BB BB AB AB BB BB AA AB 1665553 9 SNP_A- rs10511886 31826555 31.826555 0.607 BB AB BB BB AB AB AB AB BB BB 1724125 9 SNP_A- rs20583 33016572 33.016572 0.452 AA AB BB BB BB BB AA AA AA AB 1648177 9 SNP_A- rs6476493 35884737 35.884737 0.691 AA AA AA AA AA AA AA AA AA AA 1717742 9 SNP_A- rs4880042 36940301 36.940301 0.393 AA AA BB BB BB BB BB BB BB BB 1671263 9 SNP_A- rs2181139 38364977 38.364977 0.25 BB AB AA AA AA AA AB AB BB AB 1681599 9 SNP_A- rs4111409 40345280 40.34528 0.262 BB BB BB BB BB BB BB BB BB BB 1666811 9 SNP_A- rs7864775 69030853 69.030853 0.548 BB BB BB BB BB BB BB BB AA AB 1727790 9 SNP_A- rs10511972 69672094 69.672094 0.619 BB BB AA AA AA AB BB AB BB BB 1699350 9 SNP_A- rs10511984 70399849 70.399849 0.75 AA AA AA AA AA AA AA AB AA AA 1753754 9 SNP_A- rs10511999 71526051 71.526051 0.595 AA AA AA AA AA AA AA AA BB AB 1748876 9 SNP_A- rs1998372 72123726 72.123726 0.369 BB BB BB BB BB AB BB BB BB BB 1750024 9 SNP_A- rs2377524 76002013 76.002013 0.321 BB BB BB BB BB BB BB BB BB AB 1733975 9 SNP_A- rs10512079 78602073 78.602073 0.25 AA AA AA AA AB AB AA AB BB BB 1707951 9 SNP_A- rs1316823 79531349 79.531349 0.643 AB AB AA AA AA AA AA AA AA AA 1655498 9 SNP_A- rs1572147 80160841 80.160841 0.634 AA AA AB AA AB AB AA AA BB AB 1721234 9 SNP_A- rs7873639 80780459 80.780459 0.286 BB BB BB BB BB BB AA AA AA AB 1757764 9 SNP_A- rs2774635 82184146 82.184146 0.286 BB BB BB BB BB BB BB BB AA AB 1685995 9 SNP_A- rs1436932 83903397 83.903397 0.476 AB AB AA AA AA AA AA AA AA AB 1698246 9 SNP_A- rs7030902 85064645 85.064645 0.548 AA AA AB BB AB AB BB BB AA AB 1743644 9 SNP_A- rs1475524 87362117 87.362117 0.357 BB BB AB AA AB AB AA AB AA AA 1642838 9 SNP_A- rs4744114 91732136 91.732136 0.452 BB BB AB AA AB AB BB BB AA AA 1683979 9 SNP_A- rs1547201 95896039 95.896039 0.548 AA AA BB BB BB BB AB AB AB AB 1645449 9 SNP_A- rs1924001 102134812 102.134812 0.643 BB BB AA AA AA AA AB AB AA AA 1751508 9 SNP_A- rs1463983 105506339 105.506339 0.429 BB BB AB AA AB AB BB BB AB AB 1724479 9 SNP_A- rs2418076 110092906 110.092906 0.298 BB BB AA AA AA AA BB BB BB BB 1653563 9 SNP_A- rs1813202 111767658 111.767658 0.286 AB AB BB BB BB BB AB AB BB BB 1744924 9 SNP_A- rs10513222 113757379 113.757379 0.321 BB BB AB BB AB AB AA AA BB BB 1731818 9 SNP_A- rs10513267 115067920 115.06792 0.75 AA AB AA AA AA AA AA AA AA AA 1733479 9 SNP_A- rs4112759 117313823 117.313823 0.75 AA AA AA AA AA AA AA AA AA AA 1643236 9 SNP_A- rs7849366 118191918 118.191918 0.286 AA AA AB BB AB AB AB AB BB BB 1750306 9 SNP_A- rs10514837 118919482 118.919482 0.321 BB BB BB BB BB BB BB BB BB BB 1686447 9 SNP_A- rs10491529 120012279 120.012279 0.25 BB AB AB AB AB AB BB BB BB BB 1656426 9 SNP_A- rs306796 121206889 121.206889 0.631 AA AA AA AA AA AA AA AA AB AB 1677789 9 SNP_A- rs7043602 126285054 126.285054 0.738 AA AA AA AA AA AA AB AB AA AA 1705544 9 SNP_A- rs883335 129165519 129.165519 0.595 BB BB AB AB AB AB AA AA AB AB 1653355 9 SNP_A- rs2269337 130602238 130.602238 0.742 BB AB AA AA AA AA AA AA AA AA 1699424 9 SNP_A- rs2809243 132799854 132.799854 0.298 BB BB AA AA AA AA AB AB AA AA 1747024 10 SNP_A- rs1392827 1234414 1.234414 0.667 AA AA AB AB AB AB AB AB AA AA 1659685 10 SNP_A- rs4880915 1747289 1.747289 0.293 BB BB BB BB BB BB BB BB BB BB 1753764 10 SNP_A- rs9329289 2532389 2.532389 0.405 AA AA AB AB AB AB AB AB AB AB 1727231 10 SNP_A- rs2388557 3181527 3.181527 0.321 BB BB BB BB BB BB AB AB BB BB 1732637 10 SNP_A- rs1679440 4468715 4.468715 0.286 BB BB BB BB BB BB BB BB BB BB 1679829 10 SNP_A- rs946785 7041660 7.04166 0.595 AA AA AB AB AB AB BB BB AB AB 1713889 10 SNP_A- rs4385796 8539643 8.539643 0.286 BB BB BB BB BB BB BB BB AA AB 1717612 10 SNP_A- rs1762757 9449776 9.449776 0.726 AA AA AA AA AA AA BB BB AA AA 1740604 10 SNP_A- rs10508380 10003736 10.003736 0.738 AA AA AB AB AA AB BB BB AA AA 1686911 10 SNP_A- rs1041044 10644387 10.644387 0.5 BB BB AB AB BB AB AB AB BB BB 1739848 10 SNP_A- rs4750093 11829643 11.829643 0.429 BB BB BB BB BB BB BB BB BB BB 1721418 10 SNP_A- rs1108131 12537753 12.537753 0.75 AB AB AB AB AA AB BB BB AA AA 1739768 10 SNP_A- rs564166 13110955 13.110955 0.738 AB AB AA AA AA AA AA AA AA AA 1737160 10 SNP_A- rs10508465 13725194 13.725194 0.429 AA AA AB AB BB AB AB AB AA AA 1678303 10 SNP_A- rs10508473 14241057 14.241057 0.417 BB BB BB BB BB BB AB AB AA AA 1669628 10 SNP_A- rs1361588 16119457 16.119457 0.298 BB BB BB BB BB BB BB BB BB AB 1714770 10 SNP_A- rs10490962 17240369 17.240369 0.56 AB AB BB BB BB BB AB AB BB BB 1700268 10 SNP_A- rs10508555 18316688 18.316688 0.441 AB AB AB AB BB AB BB BB BB BB 1744374 10 SNP_A- rs984292 19028813 19.028813 0.393 BB BB AB AB AA AB BB BB AA AB 1748644 10 SNP_A- rs2358348 19533421 19.533421 0.643 AA AA BB BB BB BB AA AA AA AA 1686549 10 SNP_A- rs788977 21229153 21.229153 0.262 BB BB AB AB BB AB BB BB BB BB 1678189 10 SNP_A- rs1417374 23168481 23.168481 0.298 BB BB AA AA AA AA AA AA AA AB 1672001 10 SNP_A- rs2150651 24829491 24.829491 0.321 BB BB BB BB BB BB BB BB BB BB 1726471 10 SNP_A- rs10508686 25367068 25.367068 0.714 AA AA AB AB AA AB AA AA AA AA 1751938 10 SNP_A- rs4747530 25876455 25.876455 0.56 AB AB AB AB BB AB AB AB AA AA 1713661 10 SNP_A- rs10508717 26712334 26.712334 0.524 AA AA AB AB AA AB BB BB BB BB 1706402 10 SNP_A- rs1970631 28271741 28.271741 0.452 AA AA BB BB BB BB AA AA AA AB 1713649 10 SNP_A- rs703041 29265782 29.265782 0.25 BB BB AB AB BB AB BB BB BB AB 1707064 10 SNP_A- rs2776644 30294654 30.294654 0.488 AB AB AB AB BB AB BB AB BB BB 1755663 10 SNP_A- rs2490527 32711123 32.711123 0.631 BB AB AB BB BB AB AA AB AA AA 1679427 10 SNP_A- rs2269101 33546185 33.546185 0.286 AA AB AB AA AA AB BB AB BB AB 1678169 10 SNP_A- rs224750 34271036 34.271036 0.619 AA AA AA AA AA AA BB AB AA AA 1674978 10 SNP_A- rs1032408 43808849 43.808849 0.738 AA AA BB BB BB BB AA AA AA AA 1722205 10 SNP_A- rs1583421 45099157 45.099157 0.583 BB AB AA AA AA AA AA AA AA AA 1700828 10 SNP_A- rs10508908 49643864 49.643864 0.5 BB BB AA BB BB AB AA AB AB AB 1718604 10 SNP_A- rs10508929 51841987 51.841987 0.423 AA AB AA BB BB AB BB BB AA AA 1741518 10 SNP_A- rs2339628 52548976 52.548976 0.679 AA AB BB BB BB BB BB BB AB AB 1674358 10 SNP_A- rs1937666 53326630 53.32663 0.464 AB AB AA BB BB AB AA AB BB BB 1665161 10 SNP_A- rs10508976 54302305 54.302305 0.56 BB BB BB AA AB AB AB AB BB BB 1648887 10 SNP_A- rs422296 54965065 54.965065 0.714 AA AA BB AA AB AB AA AA AB AB 1660432 10 SNP_A- rs6481257 58608558 58.608558 0.274 BB BB BB BB BB BB BB BB BB BB 1642640 10 SNP_A- rs10509093 60193775 60.193775 0.452 AA AA BB BB BB BB AA AA AA AA 1697033 10 SNP_A- rs4245585 61596196 61.596196 0.286 BB BB BB AA AB AB BB BB BB BB 1723683 10 SNP_A- rs10509139 62150158 62.150158 0.691 AB AB AA AA AA AA AA AA AA AA 1653973 10 SNP_A- rs2787720 63018471 63.018471 0.488 BB BB BB BB BB BB BB BB BB BB 1713014 10 SNP_A- rs1255484 65108003 65.108003 0.488 BB BB BB BB BB BB AB AB AA AA 1667099 10 SNP_A- rs7073489 67452445 67.452445 0.274 AB AB BB BB BB BB BB BB BB BB 1658163 10 SNP_A- rs4746654 68476694 68.476694 0.441 AB AB BB BB BB BB BB BB AB AB 1642112 10 SNP_A- rs7918860 70340702 70.340702 0.583 BB BB AA BB AB AB AA AA AB AB 1720304 10 SNP_A- rs10509321 71655739 71.655739 0.298 AB AB AA AA AA AA BB BB AB AB 1729287 10 SNP_A- rs10509334 73110058 73.110058 0.427 BB BB BB BB BB BB AB AB AA AA 1707688 10 SNP_A- rs1865636 77473753 77.473753 0.5 AA AA BB AA AB AB BB BB BB BB 1654508 10 SNP_A- rs10509384 78693188 78.693188 0.726 AB AB AA AA AA AA AB AB AB AB 1697249 10 SNP_A- rs1344967 79197624 79.197624 0.262 AA AA BB BB BB BB BB BB AB AB 1679101 10 SNP_A- rs10509397 79905374 79.905374 0.476 BB BB AA BB AB AB AB AB AA AA 1748530 10 SNP_A- rs7914988 80540330 80.54033 0.441 AB AB AA BB AB AB AA AA AB AB 1736610 10 SNP_A- rs342372 84579316 84.579316 0.536 AB AB BB AA AB AB AA AA AB AB 1665139 10 SNP_A- rs2067731 86973180 86.97318 0.381 BB BB AB AA AB AB BB BB AA AA 1715818 10 SNP_A- rs2949392 87497414 87.497414 0.5 AA AA AB AA AB AB BB BB AB AB 1689101 10 SNP_A- rs391683 90510663 90.510663 0.679 BB BB AB AA AB AB AB AB AA AA 1657815 10 SNP_A- rs303212 91151335 91.151335 0.298 AB AB BB BB BB BB BB BB BB BB 1706118 10 SNP_A- rs747334 92734724 92.734724 0.476 AA AA AB AB AB AB AB AB AA AA 1703070 10 SNP_A- rs716361 93308518 93.308518 0.321 AB AB BB BB BB BB BB BB AB AB 1717632 10 SNP_A- rs2490739 94587885 94.587885 0.631 AA AA AA AA AA AA AA AA AA AA 1713435 10 SNP_A- rs3781270 95520148 95.520148 0.607 AA AA AA AA AA AA BB BB AB AB 1642080 10 SNP_A- rs10509692 97226588 97.226588 0.583 AB AB AB AB AB AB AA AA AB AB 1680183 10 SNP_A- rs10509700 97884521 97.884521 0.5 AA AA AB AB AB AB AA AA BB BB 1705694 10 SNP_A- rs793515 98978459 98.978459 0.345 BB BB BB BB BB BB BB BB BB BB 1753314 10 SNP_A- rs10509754 103711687 103.711687 0.262 AB AB BB BB BB BB BB BB AB AB 1742188 10 SNP_A- rs2451500 106622867 106.622867 0.707 AA AA AA AA AA AA AB AB BB BB 1716744 10 SNP_A- rs10509832 109070424 109.070424 0.667 AA AA BB BB BB BB BB BB AB AB 1684935 10 SNP_A- rs4113 111223756 111.223756 0.369 AA AA AB AB AB AB BB BB BB BB 1676403 10 SNP_A- rs7099088 114343455 114.343455 0.631 AB AB AB AB AB AB AB AB AA AA 1726183 10 SNP_A- rs10509976 115170888 115.170888 0.286 BB BB BB BB BB BB BB BB AB AB 1732939 10 SNP_A- rs2420070 116671318 116.671318 0.548 AB AB BB BB BB BB BB BB BB BB 1675599 10 SNP_A- rs4447088 117536799 117.536799 0.274 BB BB BB BB BB BB AB AB AB AB 1700278 10 SNP_A- rs880977 118409221 118.409221 0.452 AB AB AA AA AA AA AB AB AB AB 1660760 10 SNP_A- rs2619111 118956986 118.956986 0.702 AA AA AB AB AB AB AA AA AA AA 1747698 10 SNP_A- rs10490913 120144426 120.144426 0.537 AB AB BB BB BB BB AB AB AB AB 1682085 10 SNP_A- rs1980030 120960017 120.960017 0.393 AB AB BB BB BB BB AB AB BB BB 1731688 10 SNP_A- rs1326654 122305416 122.305416 0.405 AB AB BB BB BB BB BB BB BB BB 1641760 10 SNP_A- rs2420995 123842994 123.842994 0.393 BB BB BB BB BB BB AB AB AB AB 1741090 10 SNP_A- rs845101 125180422 125.180422 0.631 AB AB AA AA AA AA AA AA AA AA 1751948 10 SNP_A- rs1278305 127801415 127.801415 0.524 BB BB BB BB BB BB AB AB BB BB 1711689 10 SNP_A- rs10510154 128412532 128.412532 0.738 AA AA AA AA AA AA AA AA AA AA 1715610 10 SNP_A- rs2251104 130003906 130.003906 0.333 AA AA BB BB BB BB AB AB AB AB 1706112 10 SNP_A- rs1886380 130596834 130.596834 0.702 AA AA AB BB AA AB AA AA AB AB 1712012 10 SNP_A- rs4077516 133237947 133.237947 0.369 AA AB BB BB BB BB BB BB AB AB 1652639 11 SNP_A- rs2499935 5066470 5.06647 0.417 BB BB AA AB BB AB BB BB BB BB 1727870 11 SNP_A- rs2001778 5575584 5.575584 0.452 BB BB BB BB BB BB BB BB AB AB 1656094 11 SNP_A- rs10500667 6284499 6.284499 0.274 BB BB AA AB BB AB BB BB BB BB 1680969 11 SNP_A- rs2595456 6841339 6.841339 0.524 BB BB BB BB BB BB BB AB AB AB 1656388 11 SNP_A- rs3884596 7488751 7.488751 0.571 AB AB BB BB BB BB AA AB AB AB 1649885 11 SNP_A- rs3993279 10627568 10.627568 0.321 BB BB AB AB AA AB BB BB BB BB 1663461 11 SNP_A- rs10500740 11167698 11.167698 0.274 AB AB BB BB BB BB AB AB AB AB 1723239 11 SNP_A- rs1344613 12408280 12.40828 0.31 AB AB BB BB BB BB AB AB BB BB 1712474 11 SNP_A- rs1894131 15104916 15.104916 0.441 AA AB AA AA AA AA AB AB BB BB 1705810 11 SNP_A- rs2190454 17490211 17.490211 0.333 AA AB BB BB BB BB BB BB AB AB 1674594 11 SNP_A- rs211102 18003069 18.003069 0.25 BB BB AB AB AA AB BB BB AB AB 1701156 11 SNP_A- rs894556 19822510 19.82251 0.56 BB BB AB AB AB AB AA AA AB AB 1685951 11 SNP_A- rs10500886 20976742 20.976742 0.607 BB AB AA AA AA AA AA AA AA AA 1685201 11 SNP_A- rs6483807 21873219 21.873219 0.5 BB BB AA AB AB AB BB BB AA AA 1668135 11 SNP_A- rs10500927 22398998 22.398998 0.262 BB AB BB BB BB BB AB AB AB AB 1713403 11 SNP_A- rs1600958 23180524 23.180524 0.388 BB BB BB AB AB AB AA AA AA AA 1697826 11 SNP_A- rs975980 24515539 24.515539 0.441 AA AB BB BB BB BB AA AA AB AB 1753516 11 SNP_A- rs10501011 25497059 25.497059 0.417 AA AB BB BB BB BB AA AA AB AB 1652525 11 SNP_A- rs980562 30413393 30.413393 0.726 AA AB AA AB AB AB AB AB AA AA 1665219 11 SNP_A- rs1848394 30965654 30.965654 0.619 BB BB BB AB AB AB BB BB BB BB 1720570 11 SNP_A- rs10488689 31659092 31.659092 0.286 BB BB AA AA AA AA BB BB AB AB 1749112 11 SNP_A- rs1033717 33023147 33.023147 0.342 BB BB BB BB BB BB BB BB BB BB 1701102 11 SNP_A- rs2136509 34753380 34.75338 0.537 BB BB BB AB AB AB AA AA AB AB 1752850 11 SNP_A- rs10501163 36830990 36.83099 0.286 AA AA AA AA AA AA AA AA BB BB 1691121 11 SNP_A- rs992118 40016469 40.016469 0.286 BB AB AA AB AB AB AA AB BB BB 1725595 11 SNP_A- rs7102885 40922404 40.922404 0.655 AA AA AA AB AB AB BB AB BB AB 1667717 11 SNP_A- rs1531932 41781058 41.781058 0.643 AA AA BB AB AA AB AA AA AA AA 1717738 11 SNP_A- rs692726 50396846 50.396846 0.321 AA AA BB BB BB BB BB AB BB BB 1709380 11 SNP_A- rs629948 55113024 55.113024 0.643 AA AA AA BB BB AB AA AA AA AA 1752494 11 SNP_A- rs1080800 56067666 56.067666 0.381 AA AB BB AA AA AB BB AB BB BB 1697650 11 SNP_A- rs540505 56621831 56.621831 0.536 AA AB BB BB BB BB BB AB BB AB 1658985 11 SNP_A- rs612688 57333672 57.333672 0.25 BB BB BB BB BB BB AA AA BB BB 1749414 11 SNP_A- rs10501369 57870148 57.870148 0.512 AA AA AA AA AA AA BB BB AA AA 1698180 11 SNP_A- rs1941030 59982334 59.982334 0.56 BB BB AA BB BB AB AB AB AA AA 1729269 11 SNP_A- rs528736 65461684 65.461684 0.393 BB AB AB BB BB AB AA AA BB BB 1656934 11 SNP_A- rs624765 69826722 69.826722 0.714 AA AB AA AA AA AA AA AA AA AA 1738462 11 SNP_A- rs527529 74298448 74.298448 0.573 AA AA BB BB BB BB AB AB AA AA 1645461 11 SNP_A- rs1793483 76653115 76.653115 0.583 AA AA AA AA AA AA AA AA AA AB 1711405 11 SNP_A- rs3819256 77379509 77.379509 0.571 BB BB AB BB AB AB BB BB AA AB 1739334 11 SNP_A- rs7128417 77883622 77.883622 0.25 BB BB BB BB BB BB BB BB BB BB 1712184 11 SNP_A- rs483089 78543310 78.54331 0.655 AA AA AB BB AB AB AA AA AA AB 1734963 11 SNP_A- rs1569168 79525377 79.525377 0.441 AB AB AB AA AB AB AA AA BB AB 1695384 11 SNP_A- rs10501496 80598450 80.59845 0.298 AB AB AA AA AA AA AB AB AA AB 1695760 11 SNP_A- rs666649 81460553 81.460553 0.56 AB AB AB BB AB AB AB AB BB AB 1679629 11 SNP_A- rs2000922 82720260 82.72026 0.619 AA AA AA AA AA AA AB AB AA AA 1674894 11 SNP_A- rs7924334 83853909 83.853909 0.714 AA AA AA AA AA AA AA AA AA AA 1645839 11 SNP_A- rs10501586 84433725 84.433725 0.726 AA AA AA AA AA AA AB AB AA AB 1654894 11 SNP_A- rs10501612 85594787 85.594787 0.342 BB BB BB BB BB BB AB AB BB BB 1756404 11 SNP_A- rs503952 86520236 86.520236 0.268 AB AB BB BB BB BB BB BB AA AB 1722491 11 SNP_A- rs10501723 89922680 89.92268 0.5 AB AB AB AA AB AB AA AA AA AB 1740548 11 SNP_A- rs1528760 90459676 90.459676 0.738 AA AA AA AA AA AA AA AA AA AB 1741388 11 SNP_A- rs10501759 91082163 91.082163 0.537 BB BB BB BB BB BB AA AA AA AA 1755135 11 SNP_A- rs554735 91994827 91.994827 0.524 AB AB AA AA AA AA BB BB AA AB 1656446 11 SNP_A- rs2605592 92842667 92.842667 0.702 AA AA AA AA AA AA AB AB BB AB 1720756 11 SNP_A- rs609493 93708735 93.708735 0.286 BB BB BB BB BB BB AA AB BB AB 1672903 11 SNP_A- rs12627 94442268 94.442268 0.607 AA AA AB AA AB AB BB BB AA AB 1659851 11 SNP_A- rs1940201 95387950 95.38795 0.31 AB AB AB AA AB AB BB BB AB AB 1649021 11 SNP_A- rs10501859 95973889 95.973889 0.298 AB AB BB BB BB BB BB AB AB AB 1706350 11 SNP_A- rs1939713 99567868 99.567868 0.631 AA AA AB AA AB AB BB AB AA AA 1670058 11 SNP_A- rs667504 100221671 100.221671 0.738 AA AA AA AA AA AA AA AA AB AB 1712712 11 SNP_A- rs313403 102697742 102.697742 0.524 AA AA BB BB BB BB AA AB AA AA 1729283 11 SNP_A- rs260818 103425315 103.425315 0.417 BB BB BB BB BB BB BB BB AB AB 1643334 11 SNP_A- rs10502051 104808805 104.808805 0.286 BB BB BB BB BB BB BB AB BB BB 1690312 11 SNP_A- rs10502080 106341710 106.34171 0.346 BB BB BB BB BB BB BB BB BB BB 1746850 11 SNP_A- rs2640757 107936868 107.936868 0.31 AA AA BB BB BB BB BB BB BB BB 1718590 11 SNP_A- rs2298501 109571744 109.571744 0.56 AA AA BB BB BB BB BB BB BB BB 1739572 11 SNP_A- rs170486 110202174 110.202174 0.452 AB AB AA AA AA AA AA AA AA AA 1742110 11 SNP_A- rs10502152 111296905 111.296905 0.321 BB BB BB BB BB BB AA AB BB BB 1720008 11 SNP_A- rs7118530 113395335 113.395335 0.357 AA AB BB BB BB BB AA AA AA AA 1658493 11 SNP_A- rs2247060 114257194 114.257194 0.536 BB AB BB BB BB BB BB AB BB BB 1689389 11 SNP_A- rs572619 115738853 115.738853 0.619 AA AA BB BB BB BB AA AB AA AA 1652091 11 SNP_A- rs660443 116265903 116.265903 0.362 AA AA AB AB AB AB AA AA AA AA 1737192 11 SNP_A- rs1219410 121294459 121.294459 0.691 BB AB AA AA AA AA AA AA BB BB 1643985 11 SNP_A- rs872414 122170647 122.170647 0.452 AA AA AA AB AB AB BB AB BB BB 1728568 11 SNP_A- rs2078158 122950070 122.95007 0.333 BB BB AA AB AB AB BB BB BB BB 1696469 11 SNP_A- rs1940751 127447038 127.447038 0.683 AA AA AA AA AA AA AA AA BB BB 1748196 11 SNP_A- rs1368850 130433518 130.433518 0.598 AA AB AA AB AB AB BB BB AA AA 1741458 11 SNP_A- rs748807 131232636 131.232636 0.452 AA AB BB BB BB BB BB BB AB AB 1732434 12 SNP_A- rs7973282 1095178 1.095178 0.738 AA AA AA AA AA AA AA AB AA AA 1644365 12 SNP_A- rs215994 2587421 2.587421 0.274 BB BB BB BB BB BB AA AB BB BB 1716332 12 SNP_A- rs4625554 4286565 4.286565 0.298 AB AB BB BB BB BB BB BB BB BB 1708039 12 SNP_A- rs1861584 5578079 5.578079 0.702 AA AA AB AB AB AB AA AB AB AB 1727428 12 SNP_A- rs4883241 9384549 9.384549 0.369 AA AA AB AB AB AB AA AA AB AB 1708085 12 SNP_A- rs560444 9940542 9.940542 0.321 BB AB BB BB BB BB BB BB BB BB 1709352 12 SNP_A- rs1009954 11789366 11.789366 0.333 BB BB AB AB AB AB BB BB BB BB 1667917 12 SNP_A- rs10505774 13327672 13.327672 0.714 AA AB AA AA AA AA BB BB AA AA 1749536 12 SNP_A- rs10492150 14935164 14.935164 0.333 BB BB BB BB BB BB BB BB BB BB 1696855 12 SNP_A- rs4366546 18267461 18.267461 0.75 AA AA AA AA AA AA AA AA AA AA 1680095 12 SNP_A- rs10505845 19976927 19.976927 0.714 AA AA AA AA AA AA BB BB BB AB 1714486 12 SNP_A- rs4131935 20632200 20.6322 0.738 BB BB AA AA AA AA AB AB BB AB 1729086 12 SNP_A- rs2417981 21483114 21.483114 0.5 BB BB AB BB AB AB AA AA AA AA 1673313 12 SNP_A- rs3884510 24249990 24.24999 0.512 AA AA AB BB AB AB AB AB AA AA 1645425 12 SNP_A- rs10505945 24803300 24.8033 0.381 BB AB BB BB BB BB BB BB BB BB 1672243 12 SNP_A- rs10505972 25379461 25.379461 0.393 BB AB AB AA AB AB AB AB BB AB 1692085 12 SNP_A- rs9300175 27617467 27.617467 0.417 BB BB BB BB BB BB BB BB AA AB 1674778 12 SNP_A- rs148898 29606383 29.606383 0.691 AA AA AA AA AA AA AA AB AA AB 1649795 12 SNP_A- rs10506065 30342307 30.342307 0.417 BB AB BB BB BB BB BB BB BB BB 1658781 12 SNP_A- rs7979386 30966129 30.966129 0.464 AA AB AA BB BB AB AA AB AA AA 1722521 12 SNP_A- rs2593998 32333520 32.33352 0.714 AA AA AA AA AA AA AA AB AA AB 1705996 12 SNP_A- rs1905428 33450742 33.450742 0.512 AA AA AA AA AA AA BB AB BB AB 1644085 12 SNP_A- rs2389276 33989158 33.989158 0.595 AA AB BB AA AA AB BB AB BB AB 1711331 12 SNP_A- rs10506124 37305503 37.305503 0.571 BB AB AA BB BB AB AA AB AA AA 1692149 12 SNP_A- rs7969928 39561348 39.561348 0.393 AA AA BB BB BB BB BB BB BB AB 1720482 12 SNP_A- rs7309345 40585255 40.585255 0.75 BB BB AA AA AA AA AA AA AA AA 1659791 12 SNP_A- rs1369610 41755818 41.755818 0.369 AA AB AA BB AB AB BB AB AA AB 1754513 12 SNP_A- rs1506678 43535759 43.535759 0.631 AA AB AA AA AA AA AA AA AA AA 1693494 12 SNP_A- rs7310869 44951653 44.951653 0.536 AA AB BB BB BB BB AA AA BB BB 1702318 12 SNP_A- rs10506292 49031020 49.03102 0.381 BB BB BB BB BB BB BB BB BB BB 1748898 12 SNP_A- rs7968810 52445260 52.44526 0.738 AA AA AA AA AA AA AA AA AA AA 1733843 12 SNP_A- rs10506393 56672989 56.672989 0.691 AA AB BB AA AA AB BB BB BB AB 1712318 12 SNP_A- rs3913094 57197682 57.197682 0.655 AA AA AA BB BB AB AA AA AA AB 1657234 12 SNP_A- rs10506408 58834234 58.834234 0.512 AA AB AA AA AA AA BB AB AA AB 1662747 12 SNP_A- rs7308021 61145687 61.145687 0.571 BB AB AA BB BB AB AA AA BB BB 1688045 12 SNP_A- rs513203 62226007 62.226007 0.56 BB BB BB AA AA AB AB AB AB AB 1749010 12 SNP_A- rs1596727 63609374 63.609374 0.583 AA AA AA AA AA AA BB BB AA AA 1730271 12 SNP_A- rs8756 64646019 64.646019 0.595 AA AA BB BB BB BB BB BB AB AB 1716970 12 SNP_A- rs10506514 65583191 65.583191 0.298 AB AB BB BB BB BB BB BB AA AA 1721334 12 SNP_A- rs7313431 66378203 66.378203 0.631 AA AA AA AA AA AA AA AA AA AA 1646303 12 SNP_A- rs710779 68249633 68.249633 0.476 AB AB BB AA AA AB AB AB BB BB 1695666 12 SNP_A- rs2567134 69233806 69.233806 0.31 BB BB BB BB BB BB AB AB BB BB 1700862 12 SNP_A- rs7960254 70109323 70.109323 0.405 AB AB BB AA AA AB BB BB BB BB 1757570 12 SNP_A- rs10506645 70671767 70.671767 0.31 BB BB BB AA AA AB AB AB BB BB 1676631 12 SNP_A- rs7964705 72103027 72.103027 0.31 AA AA BB AA AA AB AB AB BB BB 1743470 12 SNP_A- rs1396226 73586112 73.586112 0.429 BB BB AA BB BB AB AB AB AB AB 1660536 12 SNP_A- rs1275643 74439702 74.439702 0.393 AB AB BB AA AA AB AB AB AA AA 1662713 12 SNP_A- rs310877 75889008 75.889008 0.274 BB BB BB BB BB BB BB BB AB AB 1667227 12 SNP_A- rs7315131 76389953 76.389953 0.381 AB AB AA AA AA AA AA AB AA AA 1732426 12 SNP_A- rs1796135 77491016 77.491016 0.524 BB BB BB AA AA AB AA AB AB AB 1667491 12 SNP_A- rs1244908 79104469 79.104469 0.631 AA AA BB BB BB BB AA AB AB AB 1643877 12 SNP_A- rs10506839 79948071 79.948071 0.31 AA AA BB BB BB BB AA AA AB AB 1737202 12 SNP_A- rs10506846 80609736 80.609736 0.655 AA AA AA BB BB AB AA AB AB AB 1700433 12 SNP_A- rs892540 81919943 81.919943 0.667 AB AB BB AA AA AB AA AA AA AA 1738317 12 SNP_A- rs7960510 82715458 82.715458 0.274 AB AB BB BB BB BB BB BB AB AB 1688895 12 SNP_A- rs839159 85096176 85.096176 0.726 AB AB BB BB BB BB AA AA AB AB 1675076 12 SNP_A- rs2635067 85762474 85.762474 0.571 AB AB AB AA AA AB AA AA BB BB 1663055 12 SNP_A- rs1019206 87893500 87.8935 0.333 BB BB BB BB BB BB BB BB BB BB 1727255 12 SNP_A- rs2731240 89061896 89.061896 0.31 BB BB BB BB BB BB AA AB AA AA 1657981 12 SNP_A- rs924328 91753976 91.753976 0.31 BB BB BB BB BB BB AA AA BB BB 1696519 12 SNP_A- rs4761590 93076279 93.076279 0.655 BB BB AA AA AA AA BB AB AB AB 1747554 12 SNP_A- rs759572 95985503 95.985503 0.524 BB BB AB AA AB AB AB AB BB BB 1701918 12 SNP_A- rs1394380 97055132 97.055132 0.75 AA AA AA AA AA AA AB AB AA AA 1734475 12 SNP_A- rs10492276 97699500 97.6995 0.691 AA AA AB BB AB AB AA AA AB AB 1690482 12 SNP_A- rs1718312 101743655 101.743655 0.393 BB BB BB BB BB BB BB BB BB BB 1704900 12 SNP_A- rs10507166 102367680 102.36768 0.369 BB BB BB BB BB BB BB BB BB BB 1698694 12 SNP_A- rs7954946 103966503 103.966503 0.333 BB BB AA AA AA AA AA AA AA AA 1731482 12 SNP_A- rs10507197 104564170 104.56417 0.583 AA AA AA AA AA AA BB BB AB AB 1689283 12 SNP_A- rs1444581 105816718 105.816718 0.488 BB BB AA AA AA AA AA AA BB BB 1723539 12 SNP_A- rs715447 107449185 107.449185 0.464 BB BB AB AB AB AB BB BB AB AB 1676727 12 SNP_A- rs10507234 108519202 108.519202 0.714 AB AB AA AA AA AA AB AB AA AA 1742456 12 SNP_A- rs4767550 116413870 116.41387 0.595 AA AA AB AB AB AB AA AA AB AB 1655216 12 SNP_A- rs1726392 117061645 117.061645 0.321 BB AB AB AB AB AB BB BB BB BB 1673501 12 SNP_A- rs3858710 118701913 118.701913 0.573 AA AB AA AA AA AA AA AA AB AB 1748666 12 SNP_A- rs1558062 124778387 124.778387 0.524 AA AB AB AB AB AB AA AB AA AA 1708029 12 SNP_A- rs345676 126581103 126.581103 0.643 AA AB BB BB BB BB BB AB AA AA 1716508 12 SNP_A- rs1983314 129393207 129.393207 0.333 BB AB BB BB BB BB AB AB BB BB 1664795 13 SNP_A- rs7985257 18787997 18.787997 0.524 BB BB AA AA AA AA BB AB AA AA 1652867 13 SNP_A- rs535233 20445317 20.445317 0.321 AB AB BB BB BB BB BB BB AA AB 1686943 13 SNP_A- rs2862901 23933239 23.933239 0.571 AB AB AA AA AA AA BB AB AA AA 1745741 13 SNP_A- rs10507349 25679528 25.679528 0.274 BB BB AA AA AA AB BB BB AB AB 1642868 13 SNP_A- rs1161470 28322644 28.322644 0.31 BB BB BB BB BB BB BB BB BB BB 1702806 13 SNP_A- rs213611 30348913 30.348913 0.705 AA AA AA AA AA AA AB AB AB AB 1664657 13 SNP_A- rs206079 31818618 31.818618 0.417 BB BB BB BB BB BB AB AB AB AB 1658585 13 SNP_A- rs4941700 32383381 32.383381 0.262 BB BB BB BB BB BB BB BB BB BB 1685897 13 SNP_A- rs1538001 33683068 33.683068 0.274 BB BB BB BB BB BB BB BB BB BB 1664201 13 SNP_A- rs6563348 35588714 35.588714 0.397 AA AA BB BB BB BB AB AB BB BB 1742432 13 SNP_A- rs2224655 36217500 36.2175 0.298 BB BB BB BB BB BB AB AB AA AA 1709846 13 SNP_A- rs1359214 36774982 36.774982 0.452 AB AB AA AA AA AA AB AB AB AB 1719156 13 SNP_A- rs10507466 37361657 37.361657 0.75 AA AA AA AA AA AA AB AB AB AB 1699940 13 SNP_A- rs2197879 38143188 38.143188 0.286 AB AB BB BB BB BB BB BB AA AA 1757710 13 SNP_A- rs4566029 39136387 39.136387 0.691 AA AA AA AA AA AA AB AB AA AA 1725889 13 SNP_A- rs7322754 40220977 40.220977 0.31 BB BB BB AB AB AB BB BB BB BB 1648291 13 SNP_A- rs1409075 42143450 42.14345 0.5 AB AB AB AB AB AB AA AA AA AA 1644487 13 SNP_A- rs9316020 42892291 42.892291 0.274 BB BB AB AB AB AB BB BB BB BB 1692131 13 SNP_A- rs9285153 43710570 43.71057 0.536 BB BB BB BB BB BB AB AB AB AB 1683729 13 SNP_A- rs10507544 46298746 46.298746 0.286 BB BB BB BB BB BB AB AB AB AB 1675092 13 SNP_A- rs1983805 48609971 48.609971 0.655 AA AA AA AA AA AA BB AB AA AA 1706220 13 SNP_A- rs1359613 49664241 49.664241 0.381 AA AB AA AA AA AA BB BB AB AB 1656586 13 SNP_A- rs9316513 50452286 50.452286 0.643 AA AB AA AA AA AA BB AB AA AA 1687875 13 SNP_A- rs1891948 52537146 52.537146 0.429 BB BB AA AA AA AA AA AB BB BB 1699260 13 SNP_A- rs9316642 53093973 53.093973 0.571 AA AB BB BB BB BB BB BB BB BB 1670827 13 SNP_A- rs1010947 53921670 53.92167 0.702 AA AB AA AA AA AA AA AA AA AA 1664927 13 SNP_A- rs10507599 54585799 54.585799 0.286 AA AB BB BB BB BB BB BB BB BB 1686045 13 SNP_A- rs2253408 55216697 55.216697 0.75 AA AA AB AB AB AB BB AB BB BB 1686285 13 SNP_A- rs959745 56718869 56.718869 0.274 BB BB BB BB BB BB BB AB BB BB 1668215 13 SNP_A- rs10492603 57769822 57.769822 0.679 AA AA BB BB BB BB AA AA AA AA 1706400 13 SNP_A- rs2786664 59574054 59.574054 0.417 BB BB BB BB BB BB BB BB AA AA 1695368 13 SNP_A- rs3102221 60652316 60.652316 0.488 AA AB AB AB AB AB AA AB AA AA 1690895 13 SNP_A- rs7323089 61693413 61.693413 0.56 AA AA AA AA AA AA AA AB AA AA 1731184 13 SNP_A- rs2134898 62767058 62.767058 0.25 BB BB AA AA AA AA BB AB BB BB 1697063 13 SNP_A- rs9317406 63717021 63.717021 0.417 BB AB BB BB BB BB BB BB AB AB 1707522 13 SNP_A- rs7321823 65570977 65.570977 0.679 BB BB AA AA AA AA AA AA AA AA 1645393 13 SNP_A- rs10492592 66385683 66.385683 0.714 AA AB AB AB AB AB AA AA AB AB 1726077 13 SNP_A- rs176343 68069259 68.069259 0.702 AA AB AA AA AA AA AA AA AA AA 1650731 13 SNP_A- rs2782448 68801734 68.801734 0.643 AA AA AA AA AA AA AA AA AA AA 1705084 13 SNP_A- rs3909263 71753222 71.753222 0.321 BB BB AB AB AB AB AA AA BB BB 1715042 13 SNP_A- rs10507812 72886773 72.886773 0.75 AA AA AB AB AB AB BB BB AA AA 1732673 13 SNP_A- rs9318226 73391987 73.391987 0.419 BB BB AA AA AA AA AA AA BB BB 1713643 13 SNP_A- rs9318324 74649787 74.649787 0.658 AA AA BB BB BB BB AA AA AB AB 1756880 13 SNP_A- rs10507835 75353682 75.353682 0.691 AA AA AA AA AA AA BB AB AA AA 1685215 13 SNP_A- rs1952548 76037205 76.037205 0.333 BB BB BB BB BB BB BB BB BB BB 1679595 13 SNP_A- rs7326108 77781442 77.781442 0.393 AB AB AA AA AA AA BB BB AB AB 1687191 13 SNP_A- rs3903388 78802822 78.802822 0.393 BB BB AB AB AB AB AA AA BB BB 1680379 13 SNP_A- rs1215462 79594011 79.594011 0.536 BB BB AB AB AB AB AB AB BB BB 1664955 13 SNP_A- rs1744600 80158809 80.158809 0.631 AB AB AB AB AB AB BB BB AA AA 1727874 13 SNP_A- rs10507917 80741431 80.741431 0.488 AA AA AA AA AA AA BB BB AB AB 1710116 13 SNP_A- rs9318868 81947326 81.947326 0.643 AB AB AA AA AA AA AA AA AB AB 1663633 13 SNP_A- rs9319022 83601961 83.601961 0.345 BB BB AB AB AB AB AB AB AB AB 1693530 13 SNP_A- rs1331567 84793816 84.793816 0.56 AA AA AA AA AA AA AA AA AA AA 1706422 13 SNP_A- rs995475 87558036 87.558036 0.357 AB AB BB BB BB BB AB AB BB BB 1733077 13 SNP_A- rs1113478 88908389 88.908389 0.417 AA AA AB AA AB AB AB AB AB AB 1679861 13 SNP_A- rs665530 90571947 90.571947 0.726 AA AA AA AA AA AA AB AB AA AA 1649205 13 SNP_A- rs1926489 91465990 91.46599 0.524 AB AB AA AA AA AA AA AA AB AB 1726887 13 SNP_A- rs913005 92275844 92.275844 0.607 AB AB BB BB BB BB AA AA AA AA 1714183 13 SNP_A- rs9301876 92819688 92.819688 0.56 AA AA BB BB BB BB AA AA AA AA 1701716 13 SNP_A- rs1412938 93661657 93.661657 0.726 AA AA AA AA AA AA AA AA AA AA 1681625 13 SNP_A- rs9302001 94261393 94.261393 0.274 AA AA BB BB BB BB AB AB BB BB 1648409 13 SNP_A- rs7324781 95032754 95.032754 0.405 AB AB BB BB AB AB BB BB BB BB 1741482 13 SNP_A- rs4603415 96610639 96.610639 0.631 AB AB AA AA AA AA AB AB AA AB 1661319 13 SNP_A- rs285067 97536416 97.536416 0.691 AA AB BB BB AB AB AA AA AA AA 1664929 13 SNP_A- rs1886553 98448739 98.448739 0.486 BB BB AA AA AB AB AA AA BB AB 1709292 13 SNP_A- rs2760306 99841672 99.841672 0.298 BB BB BB BB BB BB AB AB BB BB 1749428 13 SNP_A- rs10508075 101237184 101.237184 0.464 AA AB AA AA AB AB AB AB AA AA 1703098 13 SNP_A- rs1015795 102023701 102.023701 0.488 AA AB BB BB BB BB AA AA BB BB 1680317 13 SNP_A- rs279927 102539019 102.539019 0.643 AA AA AA AA AA AA AA AA BB AB 1704124 13 SNP_A- rs1033147 103460337 103.460337 0.333 AA AA BB BB BB BB AB AB BB BB 1752530 13 SNP_A- rs9300981 104440279 104.440279 0.286 BB AB BB BB BB BB AB AB BB AB 1654228 13 SNP_A- rs7318881 105459909 105.459909 0.671 BB BB AA AA AA AA BB BB BB BB 1715354 13 SNP_A- rs7327250 106243682 106.243682 0.329 BB AB BB BB BB BB BB AB BB BB 1645715 13 SNP_A- rs1320446 106965333 106.965333 0.679 BB BB BB BB AB AB BB AB AA AA 1756346 13 SNP_A- rs231604 107524007 107.524007 0.381 AA AA BB BB BB BB BB BB BB BB 1732084 13 SNP_A- rs4772985 108080882 108.080882 0.25 BB BB BB BB BB BB BB BB BB BB 1681321 13 SNP_A- rs10492480 108947427 108.947427 0.333 BB BB AA AA AA AA AA AA BB BB 1648777 13 SNP_A- rs2183850 110513987 110.513987 0.714 BB BB AA AA AA AA BB BB AA AA 1654860 14 SNP_A- rs1952805 19586195 19.586195 0.345 BB BB BB BB BB BB BB BB AB AB 1733261 14 SNP_A- rs1923 22511019 22.511019 0.429 BB AB AA AA AA AA AB AB AA AA 1702470 14 SNP_A- rs4983041 24495978 24.495978 0.595 BB AB AB AA BB AB AA AA AB AB 1645139 14 SNP_A- rs10483331 26546808 26.546808 0.417 AB AB BB BB BB BB AB AB AA AA 1676969 14 SNP_A- rs4981658 27234585 27.234585 0.732 AA AA AA AA AA AA AB AB AA AA 1680111 14 SNP_A- rs2333423 28146939 28.146939 0.381 AB AB BB BB BB BB BB BB BB BB 1734437 14 SNP_A- rs10483350 28885906 28.885906 0.738 AA AA BB BB BB BB AA AA AA AA 1669916 14 SNP_A- rs225842 29622687 29.622687 0.441 AB AB BB BB BB BB AB AB AB AB 1732697 14 SNP_A- rs1278891 31464813 31.464813 0.595 AA AA AB AA AB AB BB BB AB AB 1656700 14 SNP_A- rs9322929 33377471 33.377471 0.345 AB AB AB BB AB AB BB BB BB BB 1740154 14 SNP_A- rs799493 34621626 34.621626 0.691 AB AB AB BB AB AB AA AA AA AA 1757044 14 SNP_A- rs847498 35546428 35.546428 0.607 AA AA AA AA AA AA AA AA AA AA 1676887 14 SNP_A- rs1950361 36101975 36.101975 0.329 AB AB BB BB BB BB AB AB AB AB 1665507 14 SNP_A- rs4901596 37659956 37.659956 0.667 AB AB AA AA AA AA AB AB BB BB 1705178 14 SNP_A- rs6571869 38248210 38.24821 0.679 AA AA AB AB AB AB AB AB AA AA 1654996 14 SNP_A- rs10483511 39587714 39.587714 0.571 AB AB AB AB AB AB AB AB AB AB 1708854 14 SNP_A- rs10498360 40670723 40.670723 0.476 AB AB BB BB BB BB BB BB AB AB 1653001 14 SNP_A- rs1951874 41387217 41.387217 0.345 AA AA AB AB AB AB BB BB AA AA 1753660 14 SNP_A- rs2010338 45895631 45.895631 0.655 AA AA BB BB BB BB AA AA AA AA 1653419 14 SNP_A- rs10483573 46932227 46.932227 0.357 BB BB AA AA AA AA AA AA AB AB 1663303 14 SNP_A- rs698340 47611853 47.611853 0.585 AB AB AA AA AA AA AA AA BB BB 1664801 14 SNP_A- rs7146291 48172131 48.172131 0.274 BB BB BB BB BB BB BB BB AB AB 1722201 14 SNP_A- rs8006972 48690753 48.690753 0.31 BB BB AB AB AA AB AA AA AB AB 1650017 14 SNP_A- rs10498420 49483793 49.483793 0.476 AB AB AB AB BB AB AB AB BB BB 1720778 14 SNP_A- rs963626 50157439 50.157439 0.691 AA AA AA AA AA AA AB AB AA AA 1743320 14 SNP_A- rs1956574 51163026 51.163026 0.655 AA AA AA AA AA AA AA AA AB AB 1641756 14 SNP_A- rs7151306 52273870 52.27387 0.607 AB AB BB BB BB BB AA AA AB AB 1669704 14 SNP_A- rs877018 52892776 52.892776 0.451 BB BB AB AB AA AB AB AB AB AB 1748272 14 SNP_A- rs1382978 55788938 55.788938 0.536 AA AA BB BB BB BB AA AB AA AA 1714357 14 SNP_A- rs10483679 56503799 56.503799 0.61 BB AB AB AB AA AB AA AA AB AB 1714205 14 SNP_A- rs238376 57129665 57.129665 0.357 AA AB BB BB BB BB AA AB AB AB 1654106 14 SNP_A- rs10498488 58658876 58.658876 0.451 BB BB AB AB BB AB AA AA AA AA 1690578 14 SNP_A- rs9323353 59240800 59.2408 0.619 AA AA AB AB AA AB AA AA AA AA 1690058 14 SNP_A- rs2296274 60986931 60.986931 0.75 BB AB AA AA AA AA AA AA AB AB 1671347 14 SNP_A- rs9285590 61521469 61.521469 0.417 AA AA AA AA AA AA BB AB BB BB 1755551 14 SNP_A- rs1271582 64634456 64.634456 0.441 BB BB AA AA AA AA BB BB BB BB 1707294 14 SNP_A- rs10483805 67325404 67.325404 0.329 BB AB AB AB BB AB BB BB AB AB 1648725 14 SNP_A- rs1956528 67858721 67.858721 0.631 BB BB AA AA AA AA BB AB AB AB 1699466 14 SNP_A- rs749397 69414068 69.414068 0.262 BB BB AA AA AA AA BB BB BB BB 1682431 14 SNP_A- rs2215132 71545542 71.545542 0.262 AA AB BB BB BB BB AA AA AB AB 1674164 14 SNP_A- rs2803971 72182227 72.182227 0.713 AA AA AA AA AA AA AA AA AB AB 1672631 14 SNP_A- rs1028258 75447775 75.447775 0.726 AA AA AB AB BB AB AA AA AA AA 1648423 14 SNP_A- rs7152153 76265708 76.265708 0.286 BB BB BB BB BB BB AA AA BB BB 1700204 14 SNP_A- rs7156671 76991628 76.991628 0.7 AA AA AB AB BB AB AA AA BB BB 1701478 14 SNP_A- rs10483905 78043978 78.043978 0.429 BB BB AA AA AA AA BB BB BB BB 1706776 14 SNP_A- rs997842 78614233 78.614233 0.679 AA AA AA AA AA AA AA AB AB AB 1667157 14 SNP_A- rs2049826 79383063 79.383063 0.345 BB BB BB BB BB BB BB BB BB BB 1722889 14 SNP_A- rs2372083 81860288 81.860288 0.452 AB AB AA AA AA AA BB AB AB AB 1645339 14 SNP_A- rs2372424 83017248 83.017248 0.438 AA AA BB BB BB BB AA AA BB BB 1704748 14 SNP_A- rs8003423 83533541 83.533541 0.643 BB BB AB AB AA AB AA AA AA AA 1757272 14 SNP_A- rs1530325 84781909 84.781909 0.702 AB AB AA AA AA AA AA AB AB AB 1644835 14 SNP_A- rs10498604 86238836 86.238836 0.298 BB BB BB BB BB BB BB BB AB AB 1680733 14 SNP_A- rs8018273 86867956 86.867956 0.524 AA AA BB BB BB BB AA AA AA AA 1702796 14 SNP_A- rs429923 87481126 87.481126 0.405 BB BB AA AA AA AA AA AA BB BB 1687761 14 SNP_A- rs1742083 90256423 90.256423 0.321 AB AB AB AB AB AB AB AB AB AB 1725723 14 SNP_A- rs10498627 91041872 91.041872 0.583 AA AA BB BB BB BB AA AA AB AB 1661389 14 SNP_A- rs2148567 93244403 93.244403 0.286 BB BB BB BB BB BB AA AA BB BB 1705392 14 SNP_A- rs1456988 97557760 97.55776 0.679 AB AB AA AA AA AA AA AA BB BB 1734665 14 SNP_A- rs200331 98457298 98.457298 0.488 AB AB AA AA AA AA AB AB AA AA 1684765 14 SNP_A- rs3918051 99023837 99.023837 0.61 BB AB AA AA AA AA AB AB AA AA 1682935 14 SNP_A- rs10484072 102695759 102.695759 0.631 AA AA AB AB AB AB BB BB AB AB 1734911 14 SNP_A- rs1048257 104475429 104.475429 0.667 AA AA AB AB AB AB BB BB AB AB 1659209 15 SNP_A- rs1405186 21306806 21.306806 0.441 BB AB BB BB BB BB BB BB BB BB 1669336 15 SNP_A- rs2169637 25517776 25.517776 0.262 BB BB AA AA AA AA BB BB BB BB 1690082 15 SNP_A- rs10519635 27330404 27.330404 0.607 BB BB AB AB AB AB BB AB BB AB 1643639 15 SNP_A- rs4779462 28026287 28.026287 0.598 BB BB AB AB AB AB AA AA AA AA 1740804 15 SNP_A- rs2219507 29646927 29.646927 0.691 AA AA AA AA AA AA BB AB AA AA 1722463 15 SNP_A- rs10519737 30756749 30.756749 0.381 BB BB BB BB BB BB BB BB BB BB 1730684 15 SNP_A- rs1343900 31431890 31.43189 0.667 AA AA AB AB AB AB AA AB BB BB 1648795 15 SNP_A- rs10519956 32849133 32.849133 0.345 AB AB BB BB BB BB BB BB AA AB 1755583 15 SNP_A- rs1948650 33827340 33.82734 0.738 AA AA AA AA AA AA AB AB AA AB 1699406 15 SNP_A- rs10518868 34381353 34.381353 0.655 AA AA AB AB AB AB AB AB AA AA 1645977 15 SNP_A- rs471122 41345866 41.345866 0.3 AA AA BB BB BB BB BB BB AA AB 1665819 15 SNP_A- rs10519044 44169166 44.169166 0.381 BB BB AB AB AB AB BB BB BB BB 1673073 15 SNP_A- rs493728 48678247 48.678247 0.441 BB BB BB BB BB BB BB BB BB BB 1740676 15 SNP_A- rs1478200 51948279 51.948279 0.357 BB AB BB BB BB BB AB AB BB BB 1709940 15 SNP_A- rs2553222 52657040 52.65704 0.691 BB BB AA AA AA AA AB AB AA AB 1720532 15 SNP_A- rs4534776 55408068 55.408068 0.463 AA AA AB AB AB AB BB BB BB AB 1654466 15 SNP_A- rs1550574 56000660 56.00066 0.536 AA AA BB BB BB BB AA AA AA AB 1752856 15 SNP_A- rs2033721 58609267 58.609267 0.714 AA AB AB AB AB AB AA AA AA AB 1735597 15 SNP_A- rs3935962 59611593 59.611593 0.536 AA AB AA AA AA AA AA AA AA AB 1642536 15 SNP_A- rs10519148 60507655 60.507655 0.25 BB BB BB BB BB BB BB BB BB BB 1726387 15 SNP_A- rs2652824 61207054 61.207054 0.286 BB BB BB BB BB BB BB BB BB BB 1750348 15 SNP_A- rs10518707 65152676 65.152676 0.488 AA AB AB AB AB AB AA AA AA AA 1741266 15 SNP_A- rs305002 67928959 67.928959 0.595 AA AA AA AA AA AA BB BB BB BB 1739192 15 SNP_A- rs2128112 69973340 69.97334 0.262 BB BB BB BB BB BB BB BB BB BB 1677047 15 SNP_A- rs3898352 74308825 74.308825 0.631 BB AB AB AB AA AB BB BB AA AA 1698770 15 SNP_A- rs1446312 75199244 75.199244 0.738 AA AA AA AA AA AA AB AB AA AB 1654378 15 SNP_A- rs7163689 76352533 76.352533 0.488 AA AB AB AB AA AB AB AB BB BB 1657306 15 SNP_A- rs1001460 77291934 77.291934 0.56 AA AA AB AB BB AB AB AB AA AB 1701268 15 SNP_A- rs1320323 79128502 79.128502 0.59 AA AA AA AA AA AA AA AA AA AA 1689635 15 SNP_A- rs1846911 80255705 80.255705 0.329 AA AA BB BB BB BB AB AB BB BB 1749864 15 SNP_A- rs10520585 83462510 83.46251 0.286 BB AB AB AB AA AB BB BB BB AB 1714319 15 SNP_A- rs1961601 84030610 84.03061 0.31 BB BB AB AB AA AB BB BB BB AB 1654264 15 SNP_A- rs1122907 84708371 84.708371 0.691 AA AA AB AB BB AB AA AA AA AA 1650691 15 SNP_A- rs10520655 85887415 85.887415 0.595 BB AB AA AA AA AA AA AA AA AB 1665669 15 SNP_A- rs3817428 87216251 87.216251 0.75 AA AA AA AA AA AA AB AB AA AA 1694944 15 SNP_A- rs1079537 89675287 89.675287 0.452 AB AB AB AB AA AB BB BB BB BB 1683397 15 SNP_A- rs10520710 90688998 90.688998 0.287 BB BB BB BB BB BB BB AB BB AB 1752072 15 SNP_A- rs1989269 91611056 91.611056 0.305 BB BB BB BB BB BB BB BB BB BB 1731798 15 SNP_A- rs10520754 92760413 92.760413 0.548 AA AA AA AA AA AA AA AA AA AA 1728656 15 SNP_A- rs4321143 93957372 93.957372 0.262 BB BB BB BB BB BB BB BB BB BB 1700188 15 SNP_A- rs1551466 99344619 99.344619 0.345 AB AB AB AB AB AB BB BB AB AB 1686439 15 SNP_A- rs352716 100155950 100.15595 0.31 AA AA BB BB BB BB BB AB AA AA 1647533

The results show heterozygosity of derived phESC lines and displays changes in genotype by comparison with the related donor genotype. Portions of heterozygous segments of the donor genome became homozygous in phESC. Chromosome-chromosome number; RS ID-RS number in dbSNP database; Base pair-base pair distance as recorded by Affimetrix GeneChip; Freq A in Cauc—the frequency of A allele in Caucasian population.

In prior research, parthenogenetic activation of mouse oocytes has resulted in homozygous embryonic stem cell lines (Lin et al., Stem Cells (2003) 21:152). In human oocytes, the suppression of the second meiotic division after oocyte parthenogenetic activation and the generation of diploid embryos does not lead to the derivation of wholly homozygous hES cells.

Based on the HLA-typing results, differentiated cells derived from all phESC lines should be wholly histocompatible with the oocyte donors, making this a method to create cells of therapeutic use (Table 19).

TABLE 19 HLA-typing for phESC cell lines MHC I MHC II HLA-A HLA-B HLA-C DRB1 DQB1 DQA1 phESC-1 A*01 B*15(63) Cw*04 DRB1*12 DQB1*06 DQA1*01 A*02 B*35 Cw*0708 DRB1*13 DQB1*03 DQA1*0505 phESC-1 A*01 B*15(63) Cw*04 DRB1*12 DQB1*06 DQA1*01 donor A*02 B*35 Cw*0708 DRB1*13 DQB1*03 DQA1*0505 phESC-3, 4, 5 A*02 B*52 Cw*03 DRB1*01 DQB1*05 DQA1*0101 A*03 B*22 Cw*04 DRB1*03 DQB1*02 DQA1*05 phESC-3, 4, A*02 B*52 Cw*03 DRB1*01 DQB1*05 DQA1*0101 5 donor A*03 B*22 Cw*04 DRB1*03 DQB1*02 DQA1*05 phESC-6 A*02 B*07 Cw*04 DRB1*04 DQB1*06 DQA1*01 A*03 B*27 Cw*07 DRB1*15 DQB1*03 DQA1*03 phESC-6 A*02 B*07 Cw*04 DRB1*04 DQB1*06 DQA1*01 donor A*03 B*27 Cw*07 DRB1*15 DQB1*03 DQA1*03 phESC-7 A*01 B*38 Cw*06 DRB1*13 DQB1*06 DQA1*0106 A*02 B*57 Cw*12 DRB1*14 DQB1*06 DQA1*0103 phESC-7 A*01 B*38 Cw*06 DRB1*13 DQB1*06 DQA1*0106 donor A*02 B*57 Cw*12 DRB1*14 DQB1*06 DQA1*0103 NSF A*25 B*15(62) Cw*12 DRB1*04 DQB1*06 DQA1*01 A*32 B*18 Cw*12 DRB1*15 DQB1*03 DQA1*03

DNA-profiling of the genetic material derived from the human fibroblasts used as feeder cells revealed no contamination of the phESC cell lines with material from the human fibroblasts (Table 19).

The phESC-1 line remained undifferentiated during ten months of culture, spanning 35 passages. The other cell lines were successfully cultivated over at least 21 passages. The cells from all phESC lines formed cystic embryoid bodies in suspension culture and gave rise to derivatives of all three germ layers: ectoderm, mesoderm, and endoderm, after differentiation in vitro (FIG. 4). Approximately 5% of embryoid bodies from the phESC-1 line gave rise to beating cells five days following plating. The phESC-6 line produced pigmented epithelial-like cells (FIG. 41, K). Ectoderm differentiation is presented by positive immunocytochemical staining for neuron specific markers neurofiliment 68 (FIG. 4A), NCAM (FIG. 4B), beta III-tubulin (FIG. 4C) and the glial cell marker GFAP (FIG. 4D, M). Differentiated cells were positive for mesoderm markers including alpha-actinin (FIG. 4G) and desmin (FIG. 4J), which are muscle specific markers, and the endothelial markers PECAM-1 (FIG. 4E) and VE-Cadherin (FIG. 4F). Endoderm differentiation is presented by positive staining of differentiated derivatives for alpha-fetoprotein. These data demonstrate that phESC can be differentiated into the three germ layers that lead to all cell types of a human body.

The altered karyotype of phESC-7 may be a reason to exclude it form clinical use. Alterations of genomic imprinting in human embryos can contribute to the development of disorders linked to maternally or paternally expressed genes (Gabriel et al., Proc Natl Acad Sci USA (1998) 95:14857). In order to investigate other characteristics of the phESC lines, and to determine their suitability for use in cell therapy, imprinting analysis was performed.

Northern blots were made and screened with DNA probes SNRPN, Peg1_2, Peg1_A, H19, and GAPDH (as an internal control) as outlined above. Blotted nucleic acids were obtained from NSF, neonatal skin fibroblasts; hES, human embryonic stem cell line derived from fertilized oocytes; 1, phESC-1; 2, phESC-3, 3, phESC-4, 4, phESC-5; 5, phESC-6; 6 phESC-7. NSF RT-, hES RT-, 1 RT-are negative controls. FIG. 3 shows the results of the imprinting blot.

The maternal imprinting gene, Peg1_A shows strong binding in all of the cell lines tested. Weaker (relative to Peg1_A), but consistent binding was observed in all of the cell lines for the maternal imprinting gene H19. SNRPN shows binding predominantly in NSF, hES, phESC-4, and phESC-6. Peg1_2 shows binding predominantly in NSF, hES, phESC-1 (weaker signal), phESC-3, phESC-5, and phESC-6. GAPDH binding confirmed similar loading of RNA in all lanes.

Although the invention has been described with reference to the above examples, it will be understood that modifications and variations are encompassed within the spirit and scope of the invention. Accordingly, the invention is limited only by the following claims.

REFERENCES

-   J. Cibelli et al., Methods for Making and Using Reprogrammed Human     Somatic Cell Nuclei and Autologous and Isogenic Human Stem Cells. US     Patent Application No. 20030232430, Dec. 18, 2003. -   H. Lin et al., Multilineage Potential of Homozygous Stem Cells     Derived from Metaphase II Oocytes. Stem Cells (2003) 21:153-161. -   K. E. Vrana et al., Nonhuman Primate Parthenogenetic Stem Cells.     PNAS (2003) 100 (Suppl 1):11911-11916. -   J. P. M. Dumoulin et al., Effect of Oxygen Concentration on Human in     vitro Fertilization and Embryo Culture. Human Reproduction. (1999)     14(2):465-469. -   B. Fischer and B. D. Bavister, Oxygen Tension in the Oviduct and     Uterus of Rhesus Monkeys, Hamsters and Rabbits. J Reprod     Fertil (1993) 99:673-679. -   D. I. Kau wan and J. A. Mitchell, Intauterine Oxygen Tension during     Oestrous Cycle in the Hamster: Patterns of Change. Comp Biochem     Physiol Comp Physiol (1994) 107(4):673-678. -   F. D. Houghton et al., Oxygen Consumption and Energy Metabolism of     the Early Mouse Embryo. Mol Reprod Dev (1996) 44:476-485. -   A. Van Soom et al., Prevalence of Apoptosis and Inner Cell     Allocation in Bovine Embryos Cultured under Different Oxygen Tension     with or without Cysteine Addition. Theriogenology (2002)     57(5):1453-1465. 

1-59. (canceled)
 60. A differentiated cell derived from a stem cell obtained from a parthenogenetically activated oocyte from a human donor.
 61. The differentiated cell of claim 60, wherein the differentiated cell is histocompatible with the oocyte donor.
 62. The differentiated cell of claim 60, wherein the differentiated cell is selected from the group consisting of a neuronal cell, cardiac cell, smooth muscle cell, striated muscle cell, endothelial cell, osteoblast, oligodendrocyte, hematopoietic cell, adipose cell, stromal cell, chondrocyte, astrocyte, dendritic cell, keratinocyte, pancreatic islet, lymphoid precursor cell, mast cell, mesodermal cell, and endodermal cell.
 63. The differentiated cell of claim 62, wherein the cell expresses neurofiliment 68, NCAM, beta III-tubulin, GFAP, or a combination thereof.
 64. The differentiated cell of claim 62, wherein the cell expresses alpha-actinin, desmin, or a combination thereof.
 65. The differentiated cell of claim 62, wherein the cell expresses PECAM-1, VE-Cadherin, or a combination thereof.
 66. The differentiated cell of claim 62, wherein the cell expresses alpha-fetoprotein.
 67. The differentiated cell of claim 62, wherein the cell comprises homoplasmic mitochondrial DNA (mtDNA). 68-89. (canceled)
 90. A method of treating a subject in need thereof, comprising administering a cellular composition comprising differentiated cells, wherein the differentiated cells are derived from a stem cell obtained from a parthenogenetically activated oocyte from a human donor.
 91. The method according to claim 90, wherein the differentiated cell is selected from the group consisting of a neuronal cell, cardiac cell, smooth muscle cell, striated muscle cell, endothelial cell, osteoblast, oligodendrocyte, hematopoietic cell, adipose cell, stromal cell, chondrocyte, astrocyte, dendritic cell, keratinocyte, pancreatic islet, lymphoid precursor cell, mast cell, mesodermal cell, and endodermal cell.
 92. The method according to claim 90, wherein the subject presents a disease selected from the group consisting of Parkinson's disease, Huntington's disease, Alzheimer's disease, ALS, spinal cord defects or injuries, multiple sclerosis, muscular dystrophy, cystic fibrosis, liver disease, diabetes, heart disease, macular degeneration, cartilage defects or injuries, burns, foot ulcers, vascular disease, urinary tract disease, AIDS, and cancer.
 93. (canceled) 